Details of experiments between P49418 and Q15027
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: ACAP1
Protein B fragment: 1-740
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.33
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): 5.3
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
Measured pKd: 4.69
Experiment series 2
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: ACAP1
Protein B fragment: 534-548
Protein B site: PxxP_1
Protein B sequence: GRPRGQPPVPPKPSI
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 4
Measured pKd: 3.47
Experiment series 3
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: ACAP1
Protein B fragment: 553-567
Protein B site: PxxP_2
Protein B sequence: GSLRSKPEPPSEDLG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: -0
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
The affinity was below the detection threshold of the assay.