Details of experiments between Q14155 and Q6SPF0
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: SAMD1
Protein B fragment: 1-538
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.66
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): 7.06
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: 5.3
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: SAMD1
Protein B fragment: 102-116
Protein B site: PxxP_1
Protein B sequence: PRRGATPPAPPRAPR
Protein B construct: Bio Ado Ado-PRRGATPPAPPRAPR-CONH2
Average holdup BI: 0.17
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: 4.08
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: SAMD1
Protein B fragment: 187-201
Protein B site: PxxP_2
Protein B sequence: PPAVAPPAGPRRAPP
Protein B construct: Bio Ado Ado-PPAVAPPAGPRRAPP-CONH2
Average holdup BI: 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: 1
The affinity was below the detection threshold of the assay.
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: SAMD1
Protein B fragment: 204-218
Protein B site: PxxP_3
Protein B sequence: VAAREPPLPPPPQPP
Protein B construct: Bio Ado Ado-VAAREPPLPPPPQPP-CONH2
Average holdup BI: -0.01
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.
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: SAMD1
Protein B fragment: 424-438
Protein B site: PxxP_4
Protein B sequence: DSPSPVPLPPGKPAL
Protein B construct: Bio Ado Ado-DSPSPVPLPPGKPAL-CONH2
Average holdup BI: 0.13
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: 3.92
Experiment series 6
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SAMD1
Protein B fragment: 450-464
Protein B site: PxxP_5
Protein B sequence: PGRKEKPSDPVEWTV
Protein B construct: Bio Ado Ado-PGRKEKPSDPVEWTV-CONH2
Average holdup BI: 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: 1
The affinity was below the detection threshold of the assay.