Details of experiments between P49418 and O15056
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SYNJ2
Protein B fragment: 1-1496
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.81
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): 2.76
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: 5.64
Experiment series 2
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ2
Protein B fragment: 1098-1112
Protein B site: PxxP_1
Protein B sequence: VPNRPRPPQPPQRPP
Protein B construct: Bio Ado Ado-VPNRPRPPQPPQRPP-CONH2
Average holdup BI: 0.18
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.11
Experiment series 3
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ2
Protein B fragment: 1099-1113
Protein B site: PxxP_2
Protein B sequence: PNRPRPPQPPQRPPP
Protein B construct: Bio Ado Ado-PNRPRPPQPPQRPPP-CONH2
Average holdup BI: 0.23
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.24
Experiment series 4
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ2
Protein B fragment: 1140-1154
Protein B site: PxxP_3
Protein B sequence: QTARLLPGAPQQPPK
Protein B construct: Bio Ado Ado-QTARLLPGAPQQPPK-CONH2
Average holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ2
Protein B fragment: 1143-1157
Protein B site: PxxP_4
Protein B sequence: RLLPGAPQQPPKART
Protein B construct: Bio Ado Ado-RLLPGAPQQPPKART-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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ2
Protein B fragment: 1219-1233
Protein B site: PxxP_5
Protein B sequence: ETPQAPPLLPRRPPP
Protein B construct: Bio Ado Ado-ETPQAPPLLPRRPPP-CONH2
Average holdup BI: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.16
Experiment series 7
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ2
Protein B fragment: 1227-1241
Protein B site: PxxP_6
Protein B sequence: LPRRPPPRVPAIKKP
Protein B construct: Bio Ado Ado-LPRRPPPRVPAIKKP-CONH2
Average holdup BI: 0.42
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.66
Experiment series 8
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ2
Protein B fragment: 1324-1338
Protein B site: PxxP_7
Protein B sequence: EAPPLVPKVPPRRKK
Protein B construct: Bio Ado Ado-EAPPLVPKVPPRRKK-CONH2
Average holdup BI: 0.79
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 5.39
Experiment series 9
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SYNJ2
Protein B fragment: 1442-1456
Protein B site: PxxP_8
Protein B sequence: RSPKRDPIDPVSAGA
Protein B construct: Bio Ado Ado-RSPKRDPIDPVSAGA-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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.