Details of experiments between P49418 and Q8TEH3
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: DENND1A
Protein B fragment: 1-1009
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.45
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): 4.05
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: 4.91
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: DENND1A
Protein B fragment: 553-567
Protein B site: PxxP_1
Protein B sequence: KSTGPVPAPPDRAAS
Protein B construct: Bio Ado Ado-KSTGPVPAPPDRAAS-CONH2
Average holdup BI: -0.04
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
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: DENND1A
Protein B fragment: 942-956
Protein B site: PxxP_2
Protein B sequence: LRPGDPPLLPPRPPQ
Protein B construct: Bio Ado Ado-LRPGDPPLLPPRPPQ-CONH2
Average holdup BI: 0.11
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: 3.84