Details of experiments between the fragments Q15772~2306-2315 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: SPEG
Protein A fragment: 2306-2315
Protein A site: 14-3-3_pThr2311
Protein A modification: (TPO)2311
Protein A sequence: KARVPTVPPR
Protein A construct: KARVP(pThr)VPPR (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: 0.19
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): 2.78
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 2

Protein A protein: SPEG
Protein A fragment: 2306-2315
Protein A site: 14-3-3_pThr2311
Protein A modification: (TPO)2311
Protein A sequence: KARVPTVPPR
Protein A construct: KARVP(pThr)VPPR (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: 0.11
Holdup BI standard deviation: 0.05
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): 2.66
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.