Details of experiments between Q14155 and P38159
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: RBMX
Protein B fragment: 1-391
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): 5.27
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: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: RBMX
Protein B fragment: 161-175
Protein B site: PxxP_1
Protein B sequence: PPKRSAPSGPVRSSS
Protein B construct: Bio Ado Ado-PPKRSAPSGPVRSSS-CONH2
Average holdup BI: -0.02
Holdup BI standard deviation: 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: 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: his6-MBP-TEVsite-SH3
Protein B protein: RBMX
Protein B fragment: 90-104
Protein B site: PxxP_2
Protein B sequence: ESGRRGPPPPPRSRG
Protein B construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Average holdup BI: 0.3
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: 4.4
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: RBMX
Protein B fragment: 91-105
Protein B site: PxxP_3
Protein B sequence: SGRRGPPPPPRSRGP
Protein B construct: Bio Ado Ado-SGRRGPPPPPRSRGP-CONH2
Average holdup BI: 0.26
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: 4.32