Details of experiments between Q7KZI7 and P61981
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: MARK2
Protein A fragment: 53-62
Protein A site: 14-3-3_pThr58
Protein A modification: (TPO)58
Protein A sequence: YRLLKTIGKG
Protein A construct: YRLLK(pThr)IGKG (crude synthetic peptide)

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

Average holdup BI: 0.75
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): 8.71
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: 4.36


Experiment series 2

Protein A protein: MARK2
Protein A fragment: 626-635
Protein A site: 14-3-3_pSer631
Protein A modification: (SEP)631
Protein A sequence: GRRGASGSIF
Protein A construct: GRRGA(pSer)GSIF (crude synthetic peptide)

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

Average holdup BI: 0.4
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): 7.16
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
Measured pKd: 3.69


Experiment series 3

Protein A protein: MARK2
Protein A fragment: 53-62
Protein A site: 14-3-3_pThr58
Protein A modification: (TPO)58
Protein A sequence: YRLLKTIGKG
Protein A construct: YRLLK(pThr)IGKG (crude synthetic peptide)

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

Average holdup BI: 0.85
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): 6.17
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
Measured pKd: 4.7


Experiment series 4

Protein A protein: MARK2
Protein A fragment: 626-635
Protein A site: 14-3-3_pSer631
Protein A modification: (SEP)631
Protein A sequence: GRRGASGSIF
Protein A construct: GRRGA(pSer)GSIF (crude synthetic peptide)

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

Average holdup BI: 0.37
Holdup BI standard deviation: 0.05
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): 5.17
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
Measured pKd: 3.72


Experiment series 5

Protein A protein: MARK2
Protein A fragment: 626-635
Protein A site: 14-3-3_pSer631
Protein A modification: (SEP)631
Protein A sequence: GRRGASGSIF
Protein A construct: GRRGA(pSer)GSIF (crude synthetic peptide)

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

Average holdup BI: 0.42
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): 5.16
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: 3.74


Experiment series 6

Protein A protein: MARK2
Protein A fragment: 591-600
Protein A site: 14-3-3_pThr596
Protein A modification: (TPO)596
Protein A sequence: VSSRSTFHAG
Protein A construct: VSSRS(pThr)FHAG (crude synthetic peptide)

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

Average holdup BI: 0.81
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): 5.02
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
Measured pKd: 4.51


Experiment series 7

Protein A protein: MARK2
Protein A fragment: 53-62
Protein A site: 14-3-3_pThr58
Protein A modification: (TPO)58
Protein A sequence: YRLLKTIGKG
Protein A construct: YRLLK(pThr)IGKG (crude synthetic peptide)

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

Average holdup BI: 0.69
Holdup BI standard deviation: 0.06
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): 3.74
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: 4.21


Experiment series 8

Protein A protein: MARK2
Protein A fragment: 566-575
Protein A site: 14-3-3_pSer571
Protein A modification: (SEP)571
Protein A sequence: PVASPSAHNI
Protein A construct: PVASP(pSer)AHNI (crude synthetic peptide)

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

Average holdup BI: 0.12
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): 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
The affinity was below the detection threshold of the assay.


Experiment series 9

Protein A protein: MARK2
Protein A fragment: 566-575
Protein A site: 14-3-3_pSer571
Protein A modification: (SEP)571
Protein A sequence: PVASPSAHNI
Protein A construct: PVASP(pSer)AHNI (crude synthetic peptide)

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

Average holdup BI: 0.13
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): 2.77
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
The affinity was below the detection threshold of the assay.


Experiment series 10

Protein A protein: MARK2
Protein A fragment: 488-497
Protein A site: 14-3-3_pSer493
Protein A modification: (SEP)493
Protein A sequence: LLERASLGQA
Protein A construct: LLERA(pSer)LGQA (crude synthetic peptide)

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

Average holdup BI: 0.11
Holdup BI standard deviation: 0.04
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): 2.64
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: MARK2
Protein A fragment: 488-497
Protein A site: 14-3-3_pSer493
Protein A modification: (SEP)493
Protein A sequence: LLERASLGQA
Protein A construct: LLERA(pSer)LGQA (crude synthetic peptide)

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

Average holdup BI: 0.11
Holdup BI standard deviation: 0.05
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): 2.1
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: MARK2
Protein A fragment: 35-44
Protein A site: 14-3-3_pSer40
Protein A modification: (SEP)40
Protein A sequence: IRGRNSATSA
Protein A construct: IRGRN(pSer)ATSA (crude synthetic peptide)

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

Average holdup BI: 0.56
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): 1.92
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
Measured pKd: 3.98


Experiment series 13

Protein A protein: MARK2
Protein A fragment: 395-404
Protein A site: 14-3-3_pSer400
Protein A modification: (SEP)400
Protein A sequence: VQRSVSANPK
Protein A construct: VQRSV(pSer)ANPK (crude synthetic peptide)

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

Average holdup BI: 0.64
Holdup BI standard deviation: 0.27
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: 4.12


Experiment series 14

Protein A protein: MARK2
Protein A fragment: 35-44
Protein A site: 14-3-3_pSer40
Protein A modification: (SEP)40
Protein A sequence: IRGRNSATSA
Protein A construct: IRGRN(pSer)ATSA (crude synthetic peptide)

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

Average holdup BI: 0.91
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): 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
Measured pKd: 4.94


Experiment series 15

Protein A protein: MARK2
Protein A fragment: 488-497
Protein A site: 14-3-3_pSer493
Protein A modification: (SEP)493
Protein A sequence: LLERASLGQA
Protein A construct: LLERA(pSer)LGQA (crude synthetic peptide)

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

Average holdup BI: 0.11
Holdup BI standard deviation: 0.08
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 16

Protein A protein: MARK2
Protein A fragment: 404-413
Protein A site: 14-3-3_pSer409
Protein A modification: (SEP)409
Protein A sequence: KQRRFSDQAA
Protein A construct: KQRRF(pSer)DQAA (crude synthetic peptide)

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

Average holdup BI: 0.43
Holdup BI standard deviation: 0.32
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): 1.49
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: 3.77


Experiment series 17

Protein A protein: MARK2
Protein A fragment: 566-575
Protein A site: 14-3-3_pSer571
Protein A modification: (SEP)571
Protein A sequence: PVASPSAHNI
Protein A construct: PVASP(pSer)AHNI (crude synthetic peptide)

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

Average holdup BI: 0.07
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): 1.18
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 18

Protein A protein: MARK2
Protein A fragment: 461-470
Protein A site: 14-3-3_pThr466
Protein A modification: (TPO)466
Protein A sequence: LERKKTTPTP
Protein A construct: LERKK(pThr)TPTP (crude synthetic peptide)

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

Average holdup BI: 0.04
Holdup BI standard deviation: 0.1
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): 0.31
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: MARK2
Protein A fragment: 404-413
Protein A site: 14-3-3_pSer409
Protein A modification: (SEP)409
Protein A sequence: KQRRFSDQAA
Protein A construct: KQRRF(pSer)DQAA (crude synthetic peptide)

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

Average holdup BI: 0.22
Holdup BI standard deviation: 0.85
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): 0.16
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 20

Protein A protein: MARK2
Protein A fragment: 319-328
Protein A site: 14-3-3_pThr324
Protein A modification: (TPO)324
Protein A sequence: NEVMATYLLL
Protein A construct: NEVMA(pThr)YLLL (crude synthetic peptide)

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

Average holdup BI: -0.11
Holdup BI standard deviation: 0.72
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): 0.12
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: MARK2
Protein A fragment: 319-328
Protein A site: 14-3-3_pThr324
Protein A modification: (TPO)324
Protein A sequence: NEVMATYLLL
Protein A construct: NEVMA(pThr)YLLL (crude synthetic peptide)

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

Average holdup BI: -0.05
Holdup BI standard deviation: 0.54
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): 0.07
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: MARK2
Protein A fragment: 461-470
Protein A site: 14-3-3_pThr466
Protein A modification: (TPO)466
Protein A sequence: LERKKTTPTP
Protein A construct: LERKK(pThr)TPTP (crude synthetic peptide)

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

Average holdup BI: 0
Holdup BI standard deviation: 0.08
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): 0.05
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: MARK2
Protein A fragment: 461-470
Protein A site: 14-3-3_pThr466
Protein A modification: (TPO)466
Protein A sequence: LERKKTTPTP
Protein A construct: LERKK(pThr)TPTP (crude synthetic peptide)

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

Average holdup BI: 0
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): 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.