Details of experiments between the fragments Q9Y3M2~21-30 and P31946~1-246
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: CBY1
Protein A fragment: 21-30
Protein A site: 14-3-3_pSer26
Protein A modification: (SEP)26
Protein A sequence: LSNLHSLDRS
Protein A construct: LSNLH(pSer)LDRS (crude synthetic peptide)

Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta

Average holdup BI: -0.06
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.67
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
The affinity was below the detection threshold of the assay.


Experiment series 2

Protein A protein: CBY1
Protein A fragment: 21-30
Protein A site: 14-3-3_pSer26
Protein A modification: (SEP)26
Protein A sequence: LSNLHSLDRS
Protein A construct: LSNLH(pSer)LDRS (crude synthetic peptide)

Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta

Average holdup BI: -0.27
Holdup BI standard deviation: 0.54
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): 0.59
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.