Details of experiments between the fragments Q9Y2U5~243-252 and P61981~1-247
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: MAP3K2
Protein A fragment: 243-252
Protein A site: 14-3-3_pSer248
Protein A modification: (SEP)248
Protein A sequence: NHQEFSDYDN
Protein A construct: NHQEF(pSer)DYDN (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: -0.26
Holdup BI standard deviation: 0.38
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): 1.02
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 2

Protein A protein: MAP3K2
Protein A fragment: 243-252
Protein A site: 14-3-3_pSer248
Protein A modification: (SEP)248
Protein A sequence: NHQEFSDYDN
Protein A construct: NHQEF(pSer)DYDN (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: 0.3
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): 0.14
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.