Details of experiments between O43516 and P00519
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: WIPF1
Protein A fragment: 1-503
Protein A construct: endogenous protein (Jurkat)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.83
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): 8.61
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: 5.7
Experiment series 2
Protein A protein: WIPF1
Protein A fragment: 138-152
Protein A site: PxxP_1
Protein A sequence: AKPFSPPSGPGRFPV
Protein A construct: Bio Ado Ado-AKPFSPPSGPGRFPV-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
Measured pKd: 3.55
Experiment series 3
Protein A protein: WIPF1
Protein A fragment: 155-169
Protein A site: PxxP_2
Protein A sequence: PGHRSGPPEPQRNRM
Protein A construct: Bio Ado Ado-PGHRSGPPEPQRNRM-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.04
Holdup BI standard deviation: 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: 2023/01/18
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: WIPF1
Protein A fragment: 165-179
Protein A site: PxxP_3
Protein A sequence: QRNRMPPPRPDVGSK
Protein A construct: Bio Ado Ado-QRNRMPPPRPDVGSK-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: WIPF1
Protein A fragment: 198-212
Protein A site: PxxP_4
Protein A sequence: PHNRGSPPVPGGPRQ
Protein A construct: Bio Ado Ado-PHNRGSPPVPGGPRQ-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: WIPF1
Protein A fragment: 247-261
Protein A site: PxxP_5
Protein A sequence: SNRPPLPPTPSRALD
Protein A construct: Bio Ado Ado-SNRPPLPPTPSRALD-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: WIPF1
Protein A fragment: 260-274
Protein A site: PxxP_6
Protein A sequence: LDDKPPPPPPPVGNR
Protein A construct: Bio Ado Ado-LDDKPPPPPPPVGNR-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 8
Protein A protein: WIPF1
Protein A fragment: 304-318
Protein A site: PxxP_7
Protein A sequence: SQAPPPPPPPSRPGP
Protein A construct: Bio Ado Ado-SQAPPPPPPPSRPGP-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 9
Protein A protein: WIPF1
Protein A fragment: 326-340
Protein A site: PxxP_8
Protein A sequence: SGNDETPRLPQRNLS
Protein A construct: Bio Ado Ado-SGNDETPRLPQRNLS-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 10
Protein A protein: WIPF1
Protein A fragment: 350-364
Protein A site: PxxP_9
Protein A sequence: SPGRSGPLPPPPSER
Protein A construct: Bio Ado Ado-SPGRSGPLPPPPSER-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 11
Protein A protein: WIPF1
Protein A fragment: 359-373
Protein A site: PxxP_10
Protein A sequence: PPPSERPPPPVRDPP
Protein A construct: Bio Ado Ado-PPPSERPPPPVRDPP-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 3.6
Experiment series 12
Protein A protein: WIPF1
Protein A fragment: 372-386
Protein A site: PxxP_11
Protein A sequence: PPGRSGPLPPPPPVS
Protein A construct: Bio Ado Ado-PPGRSGPLPPPPPVS-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 13
Protein A protein: WIPF1
Protein A fragment: 390-404
Protein A site: PxxP_12
Protein A sequence: STSRALPATPQLPSR
Protein A construct: Bio Ado Ado-STSRALPATPQLPSR-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 14
Protein A protein: WIPF1
Protein A fragment: 393-407
Protein A site: PxxP_13
Protein A sequence: RALPATPQLPSRSGV
Protein A construct: Bio Ado Ado-RALPATPQLPSRSGV-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 15
Protein A protein: WIPF1
Protein A fragment: 408-422
Protein A site: PxxP_14
Protein A sequence: DSPRSGPRPPLPPDR
Protein A construct: Bio Ado Ado-DSPRSGPRPPLPPDR-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 16
Protein A protein: WIPF1
Protein A fragment: 411-425
Protein A site: PxxP_15
Protein A sequence: RSGPRPPLPPDRPSA
Protein A construct: Bio Ado Ado-RSGPRPPLPPDRPSA-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 17
Protein A protein: WIPF1
Protein A fragment: 490-503
Protein A site: PxxP_16
Protein A sequence: RRERGAPPLPPIPR_
Protein A construct: Bio Ado Ado-RRERGAPPLPPIPR_-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 3.82