Details of experiments between P49418 and Q8IZ21
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: PHACTR4
Protein B fragment: 1-702
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.11
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): 0.86
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: PHACTR4
Protein B fragment: 161-175
Protein B site: PxxP_1
Protein B sequence: ENVPKPPLLPPKRPL
Protein B construct: Bio Ado Ado-ENVPKPPLLPPKRPL-CONH2
Average holdup BI: 0.1
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: 3.79
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: PHACTR4
Protein B fragment: 229-243
Protein B site: PxxP_2
Protein B sequence: PAPRTLPAAPASTNT
Protein B construct: Bio Ado Ado-PAPRTLPAAPASTNT-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: 2023/01/16
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: PHACTR4
Protein B fragment: 253-267
Protein B site: PxxP_3
Protein B sequence: VPAKQPPIPPPKPAH
Protein B construct: Bio Ado Ado-VPAKQPPIPPPKPAH-CONH2
Average holdup BI: -0
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: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PHACTR4
Protein B fragment: 286-300
Protein B site: PxxP_4
Protein B sequence: LLSKPSPPLPPKRGI
Protein B construct: Bio Ado Ado-LLSKPSPPLPPKRGI-CONH2
Average holdup BI: 0.37
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: 2023/01/16
Number of measurements: 2
Measured pKd: 4.56
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: PHACTR4
Protein B fragment: 287-301
Protein B site: PxxP_5
Protein B sequence: LSKPSPPLPPKRGIP
Protein B construct: Bio Ado Ado-LSKPSPPLPPKRGIP-CONH2
Average holdup BI: 0.41
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: 4.64
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: PHACTR4
Protein B fragment: 346-360
Protein B site: PxxP_6
Protein B sequence: PLPTHIPPEPPRTPP
Protein B construct: Bio Ado Ado-PLPTHIPPEPPRTPP-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
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: PHACTR4
Protein B fragment: 353-367
Protein B site: PxxP_7
Protein B sequence: PEPPRTPPFPAKTFQ
Protein B construct: Bio Ado Ado-PEPPRTPPFPAKTFQ-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 9
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PHACTR4
Protein B fragment: 400-414
Protein B site: PxxP_8
Protein B sequence: DQNFGEPHIPSRLPP
Protein B construct: Bio Ado Ado-DQNFGEPHIPSRLPP-CONH2
Average holdup BI: -0.06
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.