Details of experiments between P49418 and Q96EV2
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: RBM33
Protein B fragment: 1-1170
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.29
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): 5.69
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: 4.61
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: RBM33
Protein B fragment: 477-491
Protein B site: PxxP_1
Protein B sequence: LEQRSPPPPPPPPTL
Protein B construct: Bio Ado Ado-LEQRSPPPPPPPPTL-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: 2023/01/16
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: RBM33
Protein B fragment: 521-535
Protein B site: PxxP_2
Protein B sequence: VFPRERPVRPALQPP
Protein B construct: Bio Ado Ado-VFPRERPVRPALQPP-CONH2
Average holdup BI: 0.08
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: 3.7
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: RBM33
Protein B fragment: 547-561
Protein B site: PxxP_3
Protein B sequence: GSATTRPFIPPRQPF
Protein B construct: Bio Ado Ado-GSATTRPFIPPRQPF-CONH2
Average holdup BI: 0.4
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: 4.61
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: RBM33
Protein B fragment: 706-720
Protein B site: PxxP_4
Protein B sequence: SNLRELPIAPSHVIE
Protein B construct: Bio Ado Ado-SNLRELPIAPSHVIE-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 6
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: RBM33
Protein B fragment: 757-771
Protein B site: PxxP_5
Protein B sequence: TEVKVKPASPVAQPK
Protein B construct: Bio Ado Ado-TEVKVKPASPVAQPK-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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: RBM33
Protein B fragment: 926-940
Protein B site: PxxP_6
Protein B sequence: PLHKVLPIKPADVEE
Protein B construct: Bio Ado Ado-PLHKVLPIKPADVEE-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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.