Details of experiments between Q14155 and Q5JSZ5
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: PRRC2B
Protein B fragment: 1-2229
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.94
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): 8.03
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: 6.22
Experiment series 2
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PRRC2B
Protein B fragment: 251-265
Protein B site: PxxP_1
Protein B sequence: KDPSLRPAQPVRKGA
Protein B construct: Bio Ado Ado-KDPSLRPAQPVRKGA-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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PRRC2B
Protein B fragment: 306-320
Protein B site: PxxP_2
Protein B sequence: PQLRLEPRVPFRQFQ
Protein B construct: Bio Ado Ado-PQLRLEPRVPFRQFQ-CONH2
Average holdup BI: 0.12
Holdup BI standard deviation: 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: 2023/01/17
Number of measurements: 2
Measured pKd: 3.87
Experiment series 4
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PRRC2B
Protein B fragment: 330-344
Protein B site: PxxP_3
Protein B sequence: RLGLSRPLRPLRQLV
Protein B construct: Bio Ado Ado-RLGLSRPLRPLRQLV-CONH2
Average holdup BI: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 3.63
Experiment series 5
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PRRC2B
Protein B fragment: 445-459
Protein B site: PxxP_4
Protein B sequence: VRKAPDPQPPPRKLH
Protein B construct: Bio Ado Ado-VRKAPDPQPPPRKLH-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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PRRC2B
Protein B fragment: 548-562
Protein B site: PxxP_5
Protein B sequence: QAEKEVPWSPSAEKA
Protein B construct: Bio Ado Ado-QAEKEVPWSPSAEKA-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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PRRC2B
Protein B fragment: 570-584
Protein B site: PxxP_6
Protein B sequence: AVHKGSPEFPAQETP
Protein B construct: Bio Ado Ado-AVHKGSPEFPAQETP-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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 8
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PRRC2B
Protein B fragment: 647-661
Protein B site: PxxP_7
Protein B sequence: QPVYPPPSHPQRTFY
Protein B construct: Bio Ado Ado-QPVYPPPSHPQRTFY-CONH2
Average holdup BI: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 3.73
Experiment series 9
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PRRC2B
Protein B fragment: 848-862
Protein B site: PxxP_8
Protein B sequence: QNLRCSPLEPDFVPD
Protein B construct: Bio Ado Ado-QNLRCSPLEPDFVPD-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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.