Details of experiments between P31947 and Q8TEW0
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 847-856
Protein B site: 14-3-3_pSer852
Protein B modification: (SEP)852
Protein B sequence: TRKSKSMDLG
Protein B construct: TRKSK(pSer)MDLG (crude synthetic peptide)

Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 113
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 1
Measured pKd: 4


Experiment series 2

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 884-893
Protein B site: 14-3-3_pSer889
Protein B modification: (SEP)889
Protein B sequence: LKKSSSLESL
Protein B construct: LKKSS(pSer)LESL (crude synthetic peptide)

Average holdup BI: 0.4
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.67


Experiment series 3

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 884-893
Protein B site: 14-3-3_pSer889
Protein B modification: (SEP)889
Protein B sequence: LKKSSSLESL
Protein B construct: LKKSS(pSer)LESL (crude synthetic peptide)

Average holdup BI: 0.31
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.77
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.59


Experiment series 4

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 1218-1227
Protein B site: 14-3-3_pSer1223
Protein B modification: (SEP)1223
Protein B sequence: QRQYSSLPRQ
Protein B construct: QRQYS(pSer)LPRQ (crude synthetic peptide)

Average holdup BI: 0.38
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.74


Experiment series 5

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 1218-1227
Protein B site: 14-3-3_pSer1223
Protein B modification: (SEP)1223
Protein B sequence: QRQYSSLPRQ
Protein B construct: QRQYS(pSer)LPRQ (crude synthetic peptide)

Average holdup BI: 0.5
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.85


Experiment series 6

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 293-302
Protein B site: 14-3-3_pThr298
Protein B modification: (TPO)298
Protein B sequence: ARGGRTLGLL
Protein B construct: ARGGR(pThr)LGLL (crude synthetic peptide)

Average holdup BI: 0.38
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.94
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.75


Experiment series 7

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 1218-1227
Protein B site: 14-3-3_pSer1223
Protein B modification: (SEP)1223
Protein B sequence: QRQYSSLPRQ
Protein B construct: QRQYS(pSer)LPRQ (crude synthetic peptide)

Average holdup BI: 0.4
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.94
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.78


Experiment series 8

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 293-302
Protein B site: 14-3-3_pThr298
Protein B modification: (TPO)298
Protein B sequence: ARGGRTLGLL
Protein B construct: ARGGR(pThr)LGLL (crude synthetic peptide)

Average holdup BI: 0.34
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.67
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.56


Experiment series 9

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 171-180
Protein B site: 14-3-3_pThr176
Protein B modification: (TPO)176
Protein B sequence: TRWSTTAGFL
Protein B construct: TRWST(pThr)AGFL (crude synthetic peptide)

Average holdup BI: 0.46
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.37
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.88


Experiment series 10

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 1351-1360
Protein B site: 14-3-3_pSer1356
Protein B modification: (SEP)1356
Protein B sequence: GRPFYS----
Protein B construct: TPEKGRPFY(pSer) (crude synthetic peptide)

Average holdup BI: 0.13
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.15
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 11

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 476-485
Protein B site: 14-3-3_pThr481
Protein B modification: (TPO)481
Protein B sequence: TSRDVTIGGS
Protein B construct: TSRDV(pThr)IGGS (crude synthetic peptide)

Average holdup BI: 0.03
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.92
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 12

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 1351-1360
Protein B site: 14-3-3_pSer1356
Protein B modification: (SEP)1356
Protein B sequence: GRPFYS----
Protein B construct: TPEKGRPFY(pSer) (crude synthetic peptide)

Average holdup BI: 0.15
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.85
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 13

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 884-893
Protein B site: 14-3-3_pSer889
Protein B modification: (SEP)889
Protein B sequence: LKKSSSLESL
Protein B construct: LKKSS(pSer)LESL (crude synthetic peptide)

Average holdup BI: 0.29
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.71
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.56


Experiment series 14

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 293-302
Protein B site: 14-3-3_pThr298
Protein B modification: (TPO)298
Protein B sequence: ARGGRTLGLL
Protein B construct: ARGGR(pThr)LGLL (crude synthetic peptide)

Average holdup BI: 0.17
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 15

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 1351-1360
Protein B site: 14-3-3_pSer1356
Protein B modification: (SEP)1356
Protein B sequence: GRPFYS----
Protein B construct: TPEKGRPFY(pSer) (crude synthetic peptide)

Average holdup BI: 0.13
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 16

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 1332-1341
Protein B site: 14-3-3_pSer1337
Protein B modification: (SEP)1337
Protein B sequence: VPPSPSQVAR
Protein B construct: VPPSP(pSer)QVAR (crude synthetic peptide)

Average holdup BI: 0.04
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 17

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 171-180
Protein B site: 14-3-3_pThr176
Protein B modification: (TPO)176
Protein B sequence: TRWSTTAGFL
Protein B construct: TRWST(pThr)AGFL (crude synthetic peptide)

Average holdup BI: 0.34
Holdup BI standard deviation: 0.18
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.75
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.55


Experiment series 18

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 710-719
Protein B site: 14-3-3_pSer715
Protein B modification: (SEP)715
Protein B sequence: RERRISHSLY
Protein B construct: RERRI(pSer)HSLY (crude synthetic peptide)

Average holdup BI: -0.49
Holdup BI standard deviation: 0.35
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 19

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 171-180
Protein B site: 14-3-3_pThr176
Protein B modification: (TPO)176
Protein B sequence: TRWSTTAGFL
Protein B construct: TRWST(pThr)AGFL (crude synthetic peptide)

Average holdup BI: 0.31
Holdup BI standard deviation: 0.17
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.59


Experiment series 20

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 476-485
Protein B site: 14-3-3_pThr481
Protein B modification: (TPO)481
Protein B sequence: TSRDVTIGGS
Protein B construct: TSRDV(pThr)IGGS (crude synthetic peptide)

Average holdup BI: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 21

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 822-831
Protein B site: 14-3-3_pSer827
Protein B modification: (SEP)827
Protein B sequence: GFGRQSMSEK
Protein B construct: GFGRQ(pSer)MSEK (crude synthetic peptide)

Average holdup BI: 0.07
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.17
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 22

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 968-977
Protein B site: 14-3-3_pSer973
Protein B modification: (SEP)973
Protein B sequence: DQPSHSLERQ
Protein B construct: DQPSH(pSer)LERQ (crude synthetic peptide)

Average holdup BI: -0.14
Holdup BI standard deviation: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 23

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 20-29
Protein B site: 14-3-3_pSer25
Protein B modification: (SEP)25
Protein B sequence: HMKVFSLIQQ
Protein B construct: HMKVF(pSer)LIQQ (crude synthetic peptide)

Average holdup BI: -0.46
Holdup BI standard deviation: 0.77
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.86
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 24

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 822-831
Protein B site: 14-3-3_pSer827
Protein B modification: (SEP)827
Protein B sequence: GFGRQSMSEK
Protein B construct: GFGRQ(pSer)MSEK (crude synthetic peptide)

Average holdup BI: 0.15
Holdup BI standard deviation: 0.17
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 25

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 710-719
Protein B site: 14-3-3_pSer715
Protein B modification: (SEP)715
Protein B sequence: RERRISHSLY
Protein B construct: RERRI(pSer)HSLY (crude synthetic peptide)

Average holdup BI: -0.21
Holdup BI standard deviation: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 26

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 476-485
Protein B site: 14-3-3_pThr481
Protein B modification: (TPO)481
Protein B sequence: TSRDVTIGGS
Protein B construct: TSRDV(pThr)IGGS (crude synthetic peptide)

Average holdup BI: 0.02
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 27

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 715-724
Protein B site: 14-3-3_pSer720
Protein B modification: (SEP)720
Protein B sequence: SHSLYSGIEG
Protein B construct: SHSLY(pSer)GIEG (crude synthetic peptide)

Average holdup BI: -0.08
Holdup BI standard deviation: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 28

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 196-205
Protein B site: 14-3-3_pSer201
Protein B modification: (SEP)201
Protein B sequence: DENYRSLPRD
Protein B construct: DENYR(pSer)LPRD (crude synthetic peptide)

Average holdup BI: 0.04
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 29

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 216-225
Protein B site: 14-3-3_pSer221
Protein B modification: (SEP)221
Protein B sequence: DNARSSLSAS
Protein B construct: DNARS(pSer)LSAS (crude synthetic peptide)

Average holdup BI: -0.06
Holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.49
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 30

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 216-225
Protein B site: 14-3-3_pSer221
Protein B modification: (SEP)221
Protein B sequence: DNARSSLSAS
Protein B construct: DNARS(pSer)LSAS (crude synthetic peptide)

Average holdup BI: -0.46
Holdup BI standard deviation: 1.21
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 31

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 196-205
Protein B site: 14-3-3_pSer201
Protein B modification: (SEP)201
Protein B sequence: DENYRSLPRD
Protein B construct: DENYR(pSer)LPRD (crude synthetic peptide)

Average holdup BI: 0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 32

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 1173-1182
Protein B site: 14-3-3_pSer1178
Protein B modification: (SEP)1178
Protein B sequence: RRRTYSFEQP
Protein B construct: RRRTY(pSer)FEQP (crude synthetic peptide)

Average holdup BI: -0.21
Holdup BI standard deviation: 0.53
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.41
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 33

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 715-724
Protein B site: 14-3-3_pSer720
Protein B modification: (SEP)720
Protein B sequence: SHSLYSGIEG
Protein B construct: SHSLY(pSer)GIEG (crude synthetic peptide)

Average holdup BI: 0.03
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 34

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 20-29
Protein B site: 14-3-3_pSer25
Protein B modification: (SEP)25
Protein B sequence: HMKVFSLIQQ
Protein B construct: HMKVF(pSer)LIQQ (crude synthetic peptide)

Average holdup BI: -0.16
Holdup BI standard deviation: 0.27
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.37
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 35

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 1332-1341
Protein B site: 14-3-3_pSer1337
Protein B modification: (SEP)1337
Protein B sequence: VPPSPSQVAR
Protein B construct: VPPSP(pSer)QVAR (crude synthetic peptide)

Average holdup BI: 0.02
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 36

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 828-837
Protein B site: 14-3-3_pThr833
Protein B modification: (TPO)833
Protein B sequence: MSEKRTKQFS
Protein B construct: MSEKR(pThr)KQFS (crude synthetic peptide)

Average holdup BI: 0.03
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.27
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 37

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 216-225
Protein B site: 14-3-3_pSer221
Protein B modification: (SEP)221
Protein B sequence: DNARSSLSAS
Protein B construct: DNARS(pSer)LSAS (crude synthetic peptide)

Average holdup BI: -0.05
Holdup BI standard deviation: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.25
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 38

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 139-148
Protein B site: 14-3-3_pSer144
Protein B modification: (SEP)144
Protein B sequence: HVRRSSDPAL
Protein B construct: HVRRS(pSer)DPAL (crude synthetic peptide)

Average holdup BI: 0.19
Holdup BI standard deviation: 0.46
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 39

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 822-831
Protein B site: 14-3-3_pSer827
Protein B modification: (SEP)827
Protein B sequence: GFGRQSMSEK
Protein B construct: GFGRQ(pSer)MSEK (crude synthetic peptide)

Average holdup BI: -0.1
Holdup BI standard deviation: 0.45
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 40

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 1332-1341
Protein B site: 14-3-3_pSer1337
Protein B modification: (SEP)1337
Protein B sequence: VPPSPSQVAR
Protein B construct: VPPSP(pSer)QVAR (crude synthetic peptide)

Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.17
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 41

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 876-885
Protein B site: 14-3-3_pSer881
Protein B modification: (SEP)881
Protein B sequence: RDVGPSLGLK
Protein B construct: RDVGP(pSer)LGLK (crude synthetic peptide)

Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.15
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 42

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 876-885
Protein B site: 14-3-3_pSer881
Protein B modification: (SEP)881
Protein B sequence: RDVGPSLGLK
Protein B construct: RDVGP(pSer)LGLK (crude synthetic peptide)

Average holdup BI: -0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.14
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 43

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 715-724
Protein B site: 14-3-3_pSer720
Protein B modification: (SEP)720
Protein B sequence: SHSLYSGIEG
Protein B construct: SHSLY(pSer)GIEG (crude synthetic peptide)

Average holdup BI: -0.02
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 44

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 710-719
Protein B site: 14-3-3_pSer715
Protein B modification: (SEP)715
Protein B sequence: RERRISHSLY
Protein B construct: RERRI(pSer)HSLY (crude synthetic peptide)

Average holdup BI: 0.07
Holdup BI standard deviation: 0.43
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 45

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 196-205
Protein B site: 14-3-3_pSer201
Protein B modification: (SEP)201
Protein B sequence: DENYRSLPRD
Protein B construct: DENYR(pSer)LPRD (crude synthetic peptide)

Average holdup BI: -0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 46

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 876-885
Protein B site: 14-3-3_pSer881
Protein B modification: (SEP)881
Protein B sequence: RDVGPSLGLK
Protein B construct: RDVGP(pSer)LGLK (crude synthetic peptide)

Average holdup BI: 0.01
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 47

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 968-977
Protein B site: 14-3-3_pSer973
Protein B modification: (SEP)973
Protein B sequence: DQPSHSLERQ
Protein B construct: DQPSH(pSer)LERQ (crude synthetic peptide)

Average holdup BI: 0.01
Holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 48

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 968-977
Protein B site: 14-3-3_pSer973
Protein B modification: (SEP)973
Protein B sequence: DQPSHSLERQ
Protein B construct: DQPSH(pSer)LERQ (crude synthetic peptide)

Average holdup BI: -0.01
Holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 49

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: PARD3
Protein B fragment: 139-148
Protein B site: 14-3-3_pSer144
Protein B modification: (SEP)144
Protein B sequence: HVRRSSDPAL
Protein B construct: HVRRS(pSer)DPAL (crude synthetic peptide)

Average holdup BI: 0
Holdup BI standard deviation: 0.27
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.