Details of experiments between P49418 and Q4KMP7
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: TBC1D10B
Protein B fragment: 1-808
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.39
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): 2.94
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: TBC1D10B
Protein B fragment: 15-29
Protein B site: PxxP_1
Protein B sequence: HGAPAAPSPPPRGSR
Protein B construct: Bio Ado Ado-HGAPAAPSPPPRGSR-CONH2
Average holdup BI: 0.56
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: 4.91
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: TBC1D10B
Protein B fragment: 673-687
Protein B site: PxxP_2
Protein B sequence: AGGAPSPPPPVRRAS
Protein B construct: Bio Ado Ado-AGGAPSPPPPVRRAS-CONH2
Average holdup BI: 0.06
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: 3.58