Details of experiments between P61981 and O43166
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: SIPA1L1
Protein B fragment: 1068-1077
Protein B site: 14-3-3_pSer1073
Protein B modification: (SEP)1073
Protein B sequence: WQRNASKGPH
Protein B construct: WQRNA(pSer)KGPH (crude synthetic peptide)

Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 115
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: 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: SIPA1L1
Protein B fragment: 1629-1638
Protein B site: 14-3-3_pThr1634
Protein B modification: (TPO)1634
Protein B sequence: RRPSYTLGMK
Protein B construct: RRPSY(pThr)LGMK (crude synthetic peptide)

Average holdup BI: 0.91
Holdup BI standard deviation: 0.01
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): 9.96
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: 4.92


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: SIPA1L1
Protein B fragment: 1559-1568
Protein B site: 14-3-3_pSer1564
Protein B modification: (SEP)1564
Protein B sequence: KFHALSSPQS
Protein B construct: KFHAL(pSer)SPQS (crude synthetic peptide)

Average holdup BI: 0.67
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): 8.41
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: 4.27


Experiment series 4

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: SIPA1L1
Protein B fragment: 1559-1568
Protein B site: 14-3-3_pSer1564
Protein B modification: (SEP)1564
Protein B sequence: KFHALSSPQS
Protein B construct: KFHAL(pSer)SPQS (crude synthetic peptide)

Average holdup BI: 0.76
Holdup BI standard deviation: 0.01
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): 8.37
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: 4.38


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: SIPA1L1
Protein B fragment: 1399-1408
Protein B site: 14-3-3_pSer1404
Protein B modification: (SEP)1404
Protein B sequence: KSQAGSTPLT
Protein B construct: KSQAG(pSer)TPLT (crude synthetic peptide)

Average holdup BI: 0.34
Holdup BI standard deviation: 0.01
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): 8.34
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 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: SIPA1L1
Protein B fragment: 1629-1638
Protein B site: 14-3-3_pThr1634
Protein B modification: (TPO)1634
Protein B sequence: RRPSYTLGMK
Protein B construct: RRPSY(pThr)LGMK (crude synthetic peptide)

Average holdup BI: 0.93
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): 6.98
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: 5.02


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: SIPA1L1
Protein B fragment: 1399-1408
Protein B site: 14-3-3_pSer1404
Protein B modification: (SEP)1404
Protein B sequence: KSQAGSTPLT
Protein B construct: KSQAG(pSer)TPLT (crude synthetic peptide)

Average holdup BI: 0.37
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): 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: 3.66


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: SIPA1L1
Protein B fragment: 1559-1568
Protein B site: 14-3-3_pSer1564
Protein B modification: (SEP)1564
Protein B sequence: KFHALSSPQS
Protein B construct: KFHAL(pSer)SPQS (crude synthetic peptide)

Average holdup BI: 0.77
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): 6.25
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: 4.42


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: SIPA1L1
Protein B fragment: 1399-1408
Protein B site: 14-3-3_pSer1404
Protein B modification: (SEP)1404
Protein B sequence: KSQAGSTPLT
Protein B construct: KSQAG(pSer)TPLT (crude synthetic peptide)

Average holdup BI: 0.28
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): 5.09
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: 3.55


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: SIPA1L1
Protein B fragment: 1728-1737
Protein B site: 14-3-3_pSer1733
Protein B modification: (SEP)1733
Protein B sequence: SSSKDSSPTL
Protein B construct: SSSKD(pSer)SPTL (crude synthetic peptide)

Average holdup BI: -0.08
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): 3.17
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 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: SIPA1L1
Protein B fragment: 1580-1589
Protein B site: 14-3-3_pSer1585
Protein B modification: (SEP)1585
Protein B sequence: LHRTLSDESI
Protein B construct: LHRTL(pSer)DESI (crude synthetic peptide)

Average holdup BI: 0.38
Holdup BI standard deviation: 0.18
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): 1.66
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: 3.67


Experiment series 12

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: SIPA1L1
Protein B fragment: 1728-1737
Protein B site: 14-3-3_pSer1733
Protein B modification: (SEP)1733
Protein B sequence: SSSKDSSPTL
Protein B construct: SSSKD(pSer)SPTL (crude synthetic peptide)

Average holdup BI: -0.06
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): 1.6
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
The affinity was below the detection threshold of the assay.


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: SIPA1L1
Protein B fragment: 1580-1589
Protein B site: 14-3-3_pSer1585
Protein B modification: (SEP)1585
Protein B sequence: LHRTLSDESI
Protein B construct: LHRTL(pSer)DESI (crude synthetic peptide)

Average holdup BI: 0.15
Holdup BI standard deviation: 0.11
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): 1.56
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 14

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: SIPA1L1
Protein B fragment: 203-212
Protein B site: 14-3-3_pSer208
Protein B modification: (SEP)208
Protein B sequence: HREYGSTSSI
Protein B construct: HREYG(pSer)TSSI (crude synthetic peptide)

Average holdup BI: 0.49
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): 1.56
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: 3.88


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: SIPA1L1
Protein B fragment: 250-259
Protein B site: 14-3-3_pSer255
Protein B modification: (SEP)255
Protein B sequence: GGGKGSGFSL
Protein B construct: GGGKG(pSer)GFSL (crude synthetic peptide)

Average holdup BI: 0.06
Holdup BI standard deviation: 0.05
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): 1.38
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: 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: SIPA1L1
Protein B fragment: 1185-1194
Protein B site: 14-3-3_pThr1190
Protein B modification: (TPO)1190
Protein B sequence: IDRQNTQSDI
Protein B construct: IDRQN(pThr)QSDI (crude synthetic peptide)

Average holdup BI: -0.17
Holdup BI standard deviation: 0.19
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): 1.04
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: 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: SIPA1L1
Protein B fragment: 1580-1589
Protein B site: 14-3-3_pSer1585
Protein B modification: (SEP)1585
Protein B sequence: LHRTLSDESI
Protein B construct: LHRTL(pSer)DESI (crude synthetic peptide)

Average holdup BI: 0.22
Holdup BI standard deviation: 0.23
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): 0.99
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: SIPA1L1
Protein B fragment: 1165-1174
Protein B site: 14-3-3_pSer1170
Protein B modification: (SEP)1170
Protein B sequence: TYRQKSMPEG
Protein B construct: TYRQK(pSer)MPEG (crude synthetic peptide)

Average holdup BI: -0.08
Holdup BI standard deviation: 0.15
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): 0.52
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 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: SIPA1L1
Protein B fragment: 1165-1174
Protein B site: 14-3-3_pSer1170
Protein B modification: (SEP)1170
Protein B sequence: TYRQKSMPEG
Protein B construct: TYRQK(pSer)MPEG (crude synthetic peptide)

Average holdup BI: 0.09
Holdup BI standard deviation: 0.16
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): 0.52
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 20

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: SIPA1L1
Protein B fragment: 250-259
Protein B site: 14-3-3_pSer255
Protein B modification: (SEP)255
Protein B sequence: GGGKGSGFSL
Protein B construct: GGGKG(pSer)GFSL (crude synthetic peptide)

Average holdup BI: -0.02
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): 0.47
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: 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: SIPA1L1
Protein B fragment: 203-212
Protein B site: 14-3-3_pSer208
Protein B modification: (SEP)208
Protein B sequence: HREYGSTSSI
Protein B construct: HREYG(pSer)TSSI (crude synthetic peptide)

Average holdup BI: -0.37
Holdup BI standard deviation: 0.46
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): 0.46
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: SIPA1L1
Protein B fragment: 156-165
Protein B site: 14-3-3_pSer161
Protein B modification: (SEP)161
Protein B sequence: PEAYPSSPRK
Protein B construct: PEAYP(pSer)SPRK (crude synthetic peptide)

Average holdup BI: 0.07
Holdup BI standard deviation: 0.14
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): 0.46
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: 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: SIPA1L1
Protein B fragment: 1728-1737
Protein B site: 14-3-3_pSer1733
Protein B modification: (SEP)1733
Protein B sequence: SSSKDSSPTL
Protein B construct: SSSKD(pSer)SPTL (crude synthetic peptide)

Average holdup BI: -0.02
Holdup BI standard deviation: 0.04
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): 0.38
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 24

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: SIPA1L1
Protein B fragment: 1165-1174
Protein B site: 14-3-3_pSer1170
Protein B modification: (SEP)1170
Protein B sequence: TYRQKSMPEG
Protein B construct: TYRQK(pSer)MPEG (crude synthetic peptide)

Average holdup BI: -0.04
Holdup BI standard deviation: 0.15
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): 0.23
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: 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: SIPA1L1
Protein B fragment: 203-212
Protein B site: 14-3-3_pSer208
Protein B modification: (SEP)208
Protein B sequence: HREYGSTSSI
Protein B construct: HREYG(pSer)TSSI (crude synthetic peptide)

Average holdup BI: -0.41
Holdup BI standard deviation: 1.36
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): 0.21
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
The affinity was below the detection threshold of the assay.


Experiment series 26

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: SIPA1L1
Protein B fragment: 1185-1194
Protein B site: 14-3-3_pThr1190
Protein B modification: (TPO)1190
Protein B sequence: IDRQNTQSDI
Protein B construct: IDRQN(pThr)QSDI (crude synthetic peptide)

Average holdup BI: -0.02
Holdup BI standard deviation: 0.07
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): 0.19
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 27

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: SIPA1L1
Protein B fragment: 1068-1077
Protein B site: 14-3-3_pSer1073
Protein B modification: (SEP)1073
Protein B sequence: WQRNASKGPH
Protein B construct: WQRNA(pSer)KGPH (crude synthetic peptide)

Average holdup BI: 0.17
Holdup BI standard deviation: 0.69
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): 0.16
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: 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: SIPA1L1
Protein B fragment: 156-165
Protein B site: 14-3-3_pSer161
Protein B modification: (SEP)161
Protein B sequence: PEAYPSSPRK
Protein B construct: PEAYP(pSer)SPRK (crude synthetic peptide)

Average holdup BI: 0.01
Holdup BI standard deviation: 0.09
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): 0.09
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 29

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: SIPA1L1
Protein B fragment: 1185-1194
Protein B site: 14-3-3_pThr1190
Protein B modification: (TPO)1190
Protein B sequence: IDRQNTQSDI
Protein B construct: IDRQN(pThr)QSDI (crude synthetic peptide)

Average holdup BI: 0.02
Holdup BI standard deviation: 0.17
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): 0.06
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: 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: SIPA1L1
Protein B fragment: 156-165
Protein B site: 14-3-3_pSer161
Protein B modification: (SEP)161
Protein B sequence: PEAYPSSPRK
Protein B construct: PEAYP(pSer)SPRK (crude synthetic peptide)

Average holdup BI: -0.01
Holdup BI standard deviation: 0.07
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): 0.05
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 31

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: SIPA1L1
Protein B fragment: 169-178
Protein B site: 14-3-3_pSer174
Protein B modification: (SEP)174
Protein B sequence: RIRQRSNSDI
Protein B construct: RIRQR(pSer)NSDI (crude synthetic peptide)

Average holdup BI: -0.08
Holdup BI standard deviation: 0.82
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): 0.05
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 32

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: SIPA1L1
Protein B fragment: 250-259
Protein B site: 14-3-3_pSer255
Protein B modification: (SEP)255
Protein B sequence: GGGKGSGFSL
Protein B construct: GGGKG(pSer)GFSL (crude synthetic peptide)

Average holdup BI: 0
Holdup BI standard deviation: 0.06
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): 0.02
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.