Details of experiments between Q14155 and Q96B97
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SH3KBP1
Protein B fragment: 1-665
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.01
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): 0.21
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: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.04
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.
Experiment series 3
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 3.74
Experiment series 4
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.11
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
Measured pKd: 3.83
Experiment series 5
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.