Details of experiments between Q14155 and Q96P48
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: ARAP1
Protein B fragment: 1-1450
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.74
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): 8
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: 5.45
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: ARAP1
Protein B fragment: 109-123
Protein B site: PxxP_1
Protein B sequence: EGLPAAPPIPPRRSC
Protein B construct: Bio Ado Ado-EGLPAAPPIPPRRSC-CONH2
Average holdup BI: 0.76
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: 5.34
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: ARAP1
Protein B fragment: 110-124
Protein B site: PxxP_2
Protein B sequence: GLPAAPPIPPRRSCL
Protein B construct: Bio Ado Ado-GLPAAPPIPPRRSCL-CONH2
Average holdup BI: 0.63
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: 5.05
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: ARAP1
Protein B fragment: 136-150
Protein B site: PxxP_3
Protein B sequence: APDPVLPPLPAKRHL
Protein B construct: Bio Ado Ado-APDPVLPPLPAKRHL-CONH2
Average holdup BI: 0.28
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: 4.35
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: ARAP1
Protein B fragment: 150-164
Protein B site: PxxP_4
Protein B sequence: LAELSVPPVPPRTGP
Protein B construct: Bio Ado Ado-LAELSVPPVPPRTGP-CONH2
Average holdup BI: 0.36
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: 4.54
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: ARAP1
Protein B fragment: 207-221
Protein B site: PxxP_5
Protein B sequence: SPPPCPPEIPPKPVR
Protein B construct: Bio Ado Ado-SPPPCPPEIPPKPVR-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 7
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ARAP1
Protein B fragment: 210-224
Protein B site: PxxP_6
Protein B sequence: PCPPEIPPKPVRLFP
Protein B construct: Bio Ado Ado-PCPPEIPPKPVRLFP-CONH2
Average holdup BI: 0.58
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: 4.95
Experiment series 8
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ARAP1
Protein B fragment: 233-247
Protein B site: PxxP_7
Protein B sequence: EVPEEGPGAPARVMT
Protein B construct: Bio Ado Ado-EVPEEGPGAPARVMT-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.