Details of experiments between P49418 and Q99490
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: AGAP2
Protein B fragment: 1-1192
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.61
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): 4.48
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: 5.21
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: AGAP2
Protein B fragment: 176-190
Protein B site: PxxP_1
Protein B sequence: RRLKVAPPPPAPKPC
Protein B construct: Bio Ado Ado-RRLKVAPPPPAPKPC-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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
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: AGAP2
Protein B fragment: 577-591
Protein B site: PxxP_2
Protein B sequence: AACKSLPSSPSHSAA
Protein B construct: Bio Ado Ado-AACKSLPSSPSHSAA-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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
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: AGAP2
Protein B fragment: 819-833
Protein B site: PxxP_3
Protein B sequence: PLSREPPPSPMVKKQ
Protein B construct: Bio Ado Ado-PLSREPPPSPMVKKQ-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: 2023/01/16
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: AGAP2
Protein B fragment: 91-105
Protein B site: PxxP_4
Protein B sequence: PALSPAPASPARPVS
Protein B construct: Bio Ado Ado-PALSPAPASPARPVS-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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.