Details of experiments between Q14155 and Q96J02
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: ITCH
Protein B fragment: 1-903
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.92
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): 12.95
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: 6.06
Experiment series 2
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2
Average holdup BI: 0.14
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
Measured pKd: 3.97
Experiment series 3
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2
Average holdup BI: 0.22
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
Measured pKd: 4.22
Experiment series 4
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: 0.54
Holdup BI standard deviation: 0.02
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: 2
Measured pKd: 4.86
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: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2
Average holdup BI: 0.03
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.