Details of experiments between P49418 and P42768
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: WAS
Protein B fragment: 1-502
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.76
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): 8.44
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: 5.5
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: WAS
Protein B fragment: 154-168
Protein B site: PxxP_1
Protein B sequence: GDRRQLPPPPTPANE
Protein B construct: Bio Ado Ado-GDRRQLPPPPTPANE-CONH2
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 3
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: WAS
Protein B fragment: 306-320
Protein B site: PxxP_2
Protein B sequence: EMRRQEPLPPPPPPS
Protein B construct: Bio Ado Ado-EMRRQEPLPPPPPPS-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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
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: WAS
Protein B fragment: 310-324
Protein B site: PxxP_3
Protein B sequence: QEPLPPPPPPSRGGN
Protein B construct: Bio Ado Ado-QEPLPPPPPPSRGGN-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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
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: WAS
Protein B fragment: 335-349
Protein B site: PxxP_4
Protein B sequence: NKGRSGPLPPVPLGI
Protein B construct: Bio Ado Ado-NKGRSGPLPPVPLGI-CONH2
Average holdup BI: -0.13
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: WAS
Protein B fragment: 353-367
Protein B site: PxxP_5
Protein B sequence: PPTPRGPPPPGRGGP
Protein B construct: Bio Ado Ado-PPTPRGPPPPGRGGP-CONH2
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 7
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: WAS
Protein B fragment: 361-375
Protein B site: PxxP_6
Protein B sequence: PPGRGGPPPPPPPAT
Protein B construct: Bio Ado Ado-PPGRGGPPPPPPPAT-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 8
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: WAS
Protein B fragment: 374-388
Protein B site: PxxP_7
Protein B sequence: ATGRSGPLPPPPPGA
Protein B construct: Bio Ado Ado-ATGRSGPLPPPPPGA-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.