Details of experiments between P61981 and Q5TCZ1
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 415-424
Protein B site: 14-3-3_pSer420
Protein B modification: (SEP)420
Protein B sequence: QRAQISSPNL
Protein B construct: QRAQI(pSer)SPNL (crude synthetic peptide)

Average holdup BI: 0.78
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.16
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: 4.43


Experiment series 2

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 415-424
Protein B site: 14-3-3_pSer420
Protein B modification: (SEP)420
Protein B sequence: QRAQISSPNL
Protein B construct: QRAQI(pSer)SPNL (crude synthetic peptide)

Average holdup BI: 0.74
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.93
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: 4.32


Experiment series 3

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 415-424
Protein B site: 14-3-3_pSer420
Protein B modification: (SEP)420
Protein B sequence: QRAQISSPNL
Protein B construct: QRAQI(pSer)SPNL (crude synthetic peptide)

Average holdup BI: 0.66
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.49
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
Measured pKd: 4.23


Experiment series 4

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 374-383
Protein B site: 14-3-3_pSer379
Protein B modification: (SEP)379
Protein B sequence: AKKEISLPIL
Protein B construct: AKKEI(pSer)LPIL (crude synthetic peptide)

Average holdup BI: 0.82
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.79
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: 4.62


Experiment series 5

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 374-383
Protein B site: 14-3-3_pSer379
Protein B modification: (SEP)379
Protein B sequence: AKKEISLPIL
Protein B construct: AKKEI(pSer)LPIL (crude synthetic peptide)

Average holdup BI: 0.77
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.45
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: 4.41


Experiment series 6

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 374-383
Protein B site: 14-3-3_pSer379
Protein B modification: (SEP)379
Protein B sequence: AKKEISLPIL
Protein B construct: AKKEI(pSer)LPIL (crude synthetic peptide)

Average holdup BI: 0.89
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.25
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
Measured pKd: 4.78


Experiment series 7

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 1047-1056
Protein B site: 14-3-3_pSer1052
Protein B modification: (SEP)1052
Protein B sequence: PAQRNSIPVS
Protein B construct: PAQRN(pSer)IPVS (crude synthetic peptide)

Average holdup BI: 0.13
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.85
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 8

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 1047-1056
Protein B site: 14-3-3_pSer1052
Protein B modification: (SEP)1052
Protein B sequence: PAQRNSIPVS
Protein B construct: PAQRN(pSer)IPVS (crude synthetic peptide)

Average holdup BI: 0.08
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.07
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 9

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 513-522
Protein B site: 14-3-3_pThr518
Protein B modification: (TPO)518
Protein B sequence: RTSTLTRPKV
Protein B construct: RTSTL(pThr)RPKV (crude synthetic peptide)

Average holdup BI: 0.12
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.66
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
The affinity was below the detection threshold of the assay.


Experiment series 10

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 1012-1021
Protein B site: 14-3-3_pSer1017
Protein B modification: (SEP)1017
Protein B sequence: VRRNSSFSTA
Protein B construct: VRRNS(pSer)FSTA (crude synthetic peptide)

Average holdup BI: 0.65
Holdup BI standard deviation: 0.28
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.24
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: 4.17


Experiment series 11

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 997-1006
Protein B site: 14-3-3_pSer1002
Protein B modification: (SEP)1002
Protein B sequence: LRRNESLTAT
Protein B construct: LRRNE(pSer)LTAT (crude synthetic peptide)

Average holdup BI: -0.13
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.22
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 1012-1021
Protein B site: 14-3-3_pSer1017
Protein B modification: (SEP)1017
Protein B sequence: VRRNSSFSTA
Protein B construct: VRRNS(pSer)FSTA (crude synthetic peptide)

Average holdup BI: 0.39
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.21
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: 3.68


Experiment series 13

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 251-260
Protein B site: 14-3-3_pThr256
Protein B modification: (TPO)256
Protein B sequence: LDRRWTLGGM
Protein B construct: LDRRW(pThr)LGGM (crude synthetic peptide)

Average holdup BI: 0.61
Holdup BI standard deviation: 0.24
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.21
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: 4.08


Experiment series 14

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 1047-1056
Protein B site: 14-3-3_pSer1052
Protein B modification: (SEP)1052
Protein B sequence: PAQRNSIPVS
Protein B construct: PAQRN(pSer)IPVS (crude synthetic peptide)

Average holdup BI: 0.25
Holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.8
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: 3.47


Experiment series 15

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 513-522
Protein B site: 14-3-3_pThr518
Protein B modification: (TPO)518
Protein B sequence: RTSTLTRPKV
Protein B construct: RTSTL(pThr)RPKV (crude synthetic peptide)

Average holdup BI: 0.08
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 129
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: 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 16

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 513-522
Protein B site: 14-3-3_pThr518
Protein B modification: (TPO)518
Protein B sequence: RTSTLTRPKV
Protein B construct: RTSTL(pThr)RPKV (crude synthetic peptide)

Average holdup BI: 0.09
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.22
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 997-1006
Protein B site: 14-3-3_pSer1002
Protein B modification: (SEP)1002
Protein B sequence: LRRNESLTAT
Protein B construct: LRRNE(pSer)LTAT (crude synthetic peptide)

Average holdup BI: -0.13
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.19
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 18

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 633-642
Protein B site: 14-3-3_pSer638
Protein B modification: (SEP)638
Protein B sequence: LSARGSSGDS
Protein B construct: LSARG(pSer)SGDS (crude synthetic peptide)

Average holdup BI: 0.15
Holdup BI standard deviation: 0.25
Immobilized partner concentration in holdup experiment (10-6M): 134
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 19

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 562-571
Protein B site: 14-3-3_pSer567
Protein B modification: (SEP)567
Protein B sequence: AFGFDSEPEL
Protein B construct: AFGFD(pSer)EPEL (crude synthetic peptide)

Average holdup BI: -0.2
Holdup BI standard deviation: 0.56
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.32
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 20

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 562-571
Protein B site: 14-3-3_pSer567
Protein B modification: (SEP)567
Protein B sequence: AFGFDSEPEL
Protein B construct: AFGFD(pSer)EPEL (crude synthetic peptide)

Average holdup BI: 0.18
Holdup BI standard deviation: 0.52
Immobilized partner concentration in holdup experiment (10-6M): 115
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: 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 21

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 562-571
Protein B site: 14-3-3_pSer567
Protein B modification: (SEP)567
Protein B sequence: AFGFDSEPEL
Protein B construct: AFGFD(pSer)EPEL (crude synthetic peptide)

Average holdup BI: -0.15
Holdup BI standard deviation: 0.62
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.2
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 22

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 997-1006
Protein B site: 14-3-3_pSer1002
Protein B modification: (SEP)1002
Protein B sequence: LRRNESLTAT
Protein B construct: LRRNE(pSer)LTAT (crude synthetic peptide)

Average holdup BI: -0.02
Holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 129
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: 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 23

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SH3PXD2A
Protein B fragment: 1012-1021
Protein B site: 14-3-3_pSer1017
Protein B modification: (SEP)1017
Protein B sequence: VRRNSSFSTA
Protein B construct: VRRNS(pSer)FSTA (crude synthetic peptide)

Average holdup BI: -0.03
Holdup BI standard deviation: 0.73
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.03
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.