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