Details of experiments between Q14289 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: PTK2B
Protein A fragment: 1-1009
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: 0.4
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): 1.32
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: PTK2B
Protein A fragment: 708-722
Protein A site: PxxP_1
Protein A sequence: TAFQEPPPKPSRPKY
Protein A construct: Bio Ado Ado-TAFQEPPPKPSRPKY-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 3.98
Experiment series 3
Protein A protein: PTK2B
Protein A fragment: 834-848
Protein A site: PxxP_2
Protein A sequence: YMNDKSPLTPEKEVG
Protein A construct: Bio Ado Ado-YMNDKSPLTPEKEVG-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 4
Protein A protein: PTK2B
Protein A fragment: 850-864
Protein A site: PxxP_3
Protein A sequence: LEFTGPPQKPPRLGA
Protein A construct: Bio Ado Ado-LEFTGPPQKPPRLGA-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.19
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: 4.12