Details of experiments between O15056 and Q5VST9
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SYNJ2
Protein A fragment: 1-1496
Protein A construct: endogenous protein (Jurkat)
Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.01
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): 0.05
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
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: SYNJ2
Protein A fragment: 1098-1112
Protein A site: PxxP_1
Protein A sequence: VPNRPRPPQPPQRPP
Protein A construct: Bio Ado Ado-VPNRPRPPQPPQRPP-CONH2
Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: SYNJ2
Protein A fragment: 1099-1113
Protein A site: PxxP_2
Protein A sequence: PNRPRPPQPPQRPPP
Protein A construct: Bio Ado Ado-PNRPRPPQPPQRPPP-CONH2
Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: SYNJ2
Protein A fragment: 1140-1154
Protein A site: PxxP_3
Protein A sequence: QTARLLPGAPQQPPK
Protein A construct: Bio Ado Ado-QTARLLPGAPQQPPK-CONH2
Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: SYNJ2
Protein A fragment: 1143-1157
Protein A site: PxxP_4
Protein A sequence: RLLPGAPQQPPKART
Protein A construct: Bio Ado Ado-RLLPGAPQQPPKART-CONH2
Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: SYNJ2
Protein A fragment: 1219-1233
Protein A site: PxxP_5
Protein A sequence: ETPQAPPLLPRRPPP
Protein A construct: Bio Ado Ado-ETPQAPPLLPRRPPP-CONH2
Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 3.57
Experiment series 7
Protein A protein: SYNJ2
Protein A fragment: 1227-1241
Protein A site: PxxP_6
Protein A sequence: LPRRPPPRVPAIKKP
Protein A construct: Bio Ado Ado-LPRRPPPRVPAIKKP-CONH2
Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 8
Protein A protein: SYNJ2
Protein A fragment: 1324-1338
Protein A site: PxxP_7
Protein A sequence: EAPPLVPKVPPRRKK
Protein A construct: Bio Ado Ado-EAPPLVPKVPPRRKK-CONH2
Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.06
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 9
Protein A protein: SYNJ2
Protein A fragment: 1442-1456
Protein A site: PxxP_8
Protein A sequence: RSPKRDPIDPVSAGA
Protein A construct: Bio Ado Ado-RSPKRDPIDPVSAGA-CONH2
Protein B protein: OBSCN
Protein B fragment: 5594-5674
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 3.54