Details of experiments between Q96T37 and Q14155
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: RBM15
Protein A fragment: 1-977
Protein A construct: endogenous protein (Jurkat)
Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.36
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): 7.87
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: 4.74
Experiment series 2
Protein A protein: RBM15
Protein A fragment: 311-325
Protein A site: PxxP_1
Protein A sequence: ALGRLPPPPPPPLPR
Protein A construct: Bio Ado Ado-ALGRLPPPPPPPLPR-CONH2
Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.08
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: 3.68
Experiment series 3
Protein A protein: RBM15
Protein A fragment: 583-597
Protein A site: PxxP_2
Protein A sequence: DSDWVPPPPPVRERS
Protein A construct: Bio Ado Ado-DSDWVPPPPPVRERS-CONH2
Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.32
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: 4.45