Details of experiments between Q96HA1 and P49418
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: POM121
Protein A fragment: 1-1249
Protein A construct: endogenous protein (Jurkat)
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.41
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): 2.4
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: POM121
Protein A fragment: 140-154
Protein A site: PxxP_1
Protein A sequence: YLGKPGPPQPAAAPE
Protein A construct: Bio Ado Ado-YLGKPGPPQPAAAPE-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: POM121
Protein A fragment: 161-175
Protein A site: PxxP_2
Protein A sequence: RPGRRPPARPAPRSP
Protein A construct: Bio Ado Ado-RPGRRPPARPAPRSP-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/16
Number of measurements: 1
Measured pKd: 4.54
Experiment series 4
Protein A protein: POM121
Protein A fragment: 166-180
Protein A site: PxxP_3
Protein A sequence: PPARPAPRSPPPRSP
Protein A construct: Bio Ado Ado-PPARPAPRSPPPRSP-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.12
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: 3.9
Experiment series 5
Protein A protein: POM121
Protein A fragment: 180-194
Protein A site: PxxP_4
Protein A sequence: PPPRSPPPSPPTHRA
Protein A construct: Bio Ado Ado-PPPRSPPPSPPTHRA-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.04
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: POM121
Protein A fragment: 442-456
Protein A site: PxxP_5
Protein A sequence: SSRSQTPERPAKKIR
Protein A construct: Bio Ado Ado-SSRSQTPERPAKKIR-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.11
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: 3.85
Experiment series 7
Protein A protein: POM121
Protein A fragment: 737-751
Protein A site: PxxP_6
Protein A sequence: KSEKEGPTPPGPSVT
Protein A construct: Bio Ado Ado-KSEKEGPTPPGPSVT-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.