Details of experiments between P49418 and Q8WXE0
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: CASKIN2
Protein B fragment: 1-1202
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.65
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): 5.09
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: 5.26
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: CASKIN2
Protein B fragment: 1121-1135
Protein B site: PxxP_1
Protein B sequence: PRLGPRPVPPPRPES
Protein B construct: Bio Ado Ado-PRLGPRPVPPPRPES-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: 4.62
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: CASKIN2
Protein B fragment: 683-697
Protein B site: PxxP_2
Protein B sequence: GGPEPLPLPPARSPS
Protein B construct: Bio Ado Ado-GGPEPLPLPPARSPS-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: 3.81
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: CASKIN2
Protein B fragment: 782-796
Protein B site: PxxP_3
Protein B sequence: AGPPATPPDPPRPKR
Protein B construct: Bio Ado Ado-AGPPATPPDPPRPKR-CONH2
Average holdup BI: 0.31
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: 4.42
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: CASKIN2
Protein B fragment: 785-799
Protein B site: PxxP_4
Protein B sequence: PATPPDPPRPKRRSH
Protein B construct: Bio Ado Ado-PATPPDPPRPKRRSH-CONH2
Average holdup BI: 0.43
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: 4.66
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: CASKIN2
Protein B fragment: 862-876
Protein B site: PxxP_5
Protein B sequence: RARRKGPPPPPPKRL
Protein B construct: Bio Ado Ado-RARRKGPPPPPPKRL-CONH2
Average holdup BI: 0.35
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: 4.51
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: CASKIN2
Protein B fragment: 863-877
Protein B site: PxxP_6
Protein B sequence: ARRKGPPPPPPKRLS
Protein B construct: Bio Ado Ado-ARRKGPPPPPPKRLS-CONH2
Average holdup BI: 0.23
Holdup BI standard deviation: 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: 2
Measured pKd: 4.24
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: CASKIN2
Protein B fragment: 864-878
Protein B site: PxxP_7
Protein B sequence: RRKGPPPPPPKRLSS
Protein B construct: Bio Ado Ado-RRKGPPPPPPKRLSS-CONH2
Average holdup BI: 0.24
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: 4.27
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: CASKIN2
Protein B fragment: 960-974
Protein B site: PxxP_8
Protein B sequence: IKQRPKPAGPPPRET
Protein B construct: Bio Ado Ado-IKQRPKPAGPPPRET-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 10
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: CASKIN2
Protein B fragment: 969-983
Protein B site: PxxP_9
Protein B sequence: PPPRETPVPPGLDFN
Protein B construct: Bio Ado Ado-PPPRETPVPPGLDFN-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.