Details of experiments between Q96J02 and P55345
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ITCH
Protein A fragment: 1-903
Protein A construct: endogenous protein (Jurkat)
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.12
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): 2.73
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: ITCH
Protein A fragment: 244-258
Protein A site: PxxP_1
Protein A sequence: SNGGFKPSRPPRPSR
Protein A construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: ITCH
Protein A fragment: 247-261
Protein A site: PxxP_2
Protein A sequence: GFKPSRPPRPSRPPP
Protein A construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
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: ITCH
Protein A fragment: 255-269
Protein A site: PxxP_3
Protein A sequence: RPSRPPPPTPRRPAS
Protein A construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.04
Holdup BI standard deviation: 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: ITCH
Protein A fragment: 353-367
Protein A site: PxxP_4
Protein A sequence: TWDRPEPLPPGWERR
Protein A construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
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.