Details of experiments between P49418 and Q9Y5K6
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: CD2AP
Protein B fragment: 1-639
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.04
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): 0.57
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: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: CD2AP
Protein B fragment: 335-349
Protein B site: PxxP_1
Protein B sequence: FPKPKKPPPPAKAPA
Protein B construct: Bio Ado Ado-FPKPKKPPPPAKAPA-CONH2
Average holdup BI: -0.05
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: CD2AP
Protein B fragment: 379-393
Protein B site: PxxP_2
Protein B sequence: PSKPAAPQVPPKKPT
Protein B construct: Bio Ado Ado-PSKPAAPQVPPKKPT-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 4
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: CD2AP
Protein B fragment: 386-400
Protein B site: PxxP_3
Protein B sequence: QVPPKKPTPPTKASN
Protein B construct: Bio Ado Ado-QVPPKKPTPPTKASN-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 5
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: CD2AP
Protein B fragment: 404-418
Protein B site: PxxP_4
Protein B sequence: SSGTVYPKRPEKPVP
Protein B construct: Bio Ado Ado-SSGTVYPKRPEKPVP-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: CD2AP
Protein B fragment: 412-426
Protein B site: PxxP_5
Protein B sequence: RPEKPVPPPPPIAKI
Protein B construct: Bio Ado Ado-RPEKPVPPPPPIAKI-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 7
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: CD2AP
Protein B fragment: 488-502
Protein B site: PxxP_6
Protein B sequence: HLTANRPKMPGRRLP
Protein B construct: Bio Ado Ado-HLTANRPKMPGRRLP-CONH2
Average holdup BI: 0.21
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: 4.19