Details of experiments between P49418 and O14733
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3

Protein B protein: MAP2K7
Protein B fragment: 1-419
Protein B construct: endogenous protein (Jurkat)

Average holdup BI: 0.25
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.23
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.29.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
Number of measurements: 3
The affinity was below the detection threshold of the assay.

Experiment series 2

Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MAP2K7
Protein B fragment: 285-299
Protein B site: PxxP_1
Protein B sequence: APERIDPPDPTKPDY
Protein B construct: Bio Ado Ado-APERIDPPDPTKPDY-CONH2

Average holdup BI: 0.01
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 2
The affinity was below the detection threshold of the assay.

Experiment series 3

Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: MAP2K7
Protein B fragment: 59-73
Protein B site: PxxP_2
Protein B sequence: ESSPQHPTPPARPRH
Protein B construct: Bio Ado Ado-ESSPQHPTPPARPRH-CONH2

Average holdup BI: 0.3
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.41