Details of experiments between Q96BH1 and P00519
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: RNF25
Protein A fragment: 1-459
Protein A construct: endogenous protein (Jurkat)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.09
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): 0.32
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: RNF25
Protein A fragment: 333-347
Protein A site: PxxP_1
Protein A sequence: QKAMLDPPKPSRGPW
Protein A construct: Bio Ado Ado-QKAMLDPPKPSRGPW-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 3.63
Experiment series 3
Protein A protein: RNF25
Protein A fragment: 369-383
Protein A site: PxxP_2
Protein A sequence: TQELPPPEGPLKEPM
Protein A construct: Bio Ado Ado-TQELPPPEGPLKEPM-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.