Details of experiments between Q9ULL5 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: PRR12
Protein A fragment: 1-2036
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: 0.89
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): 6.47
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: 5.92
Experiment series 2
Protein A protein: PRR12
Protein A fragment: 1215-1229
Protein A site: PxxP_1
Protein A sequence: GGTRLEPLKPLKIKL
Protein A construct: Bio Ado Ado-GGTRLEPLKPLKIKL-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.02
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 3
Protein A protein: PRR12
Protein A fragment: 13-27
Protein A site: PxxP_2
Protein A modification: BAA86519.1
Protein A sequence: HPVRPPPPPPPPMPL
Protein A construct: Bio Ado Ado-HPVRPPPPPPPPMPL-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 4
Protein A protein: PRR12
Protein A fragment: 1300-1314
Protein A site: PxxP_3
Protein A sequence: QAGLTPPLSPPKSVP
Protein A construct: Bio Ado Ado-QAGLTPPLSPPKSVP-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 5
Protein A protein: PRR12
Protein A fragment: 1453-1467
Protein A site: PxxP_4
Protein A sequence: LEEKPPPTPPPAPTP
Protein A construct: Bio Ado Ado-LEEKPPPTPPPAPTP-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: PRR12
Protein A fragment: 1664-1678
Protein A site: PxxP_5
Protein A sequence: RKPWHRPPVPVRRSG
Protein A construct: Bio Ado Ado-RKPWHRPPVPVRRSG-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.09
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: 3.74
Experiment series 7
Protein A protein: PRR12
Protein A fragment: 1691-1705
Protein A site: PxxP_6
Protein A sequence: APPPKAPAPPPKPET
Protein A construct: Bio Ado Ado-APPPKAPAPPPKPET-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 8
Protein A protein: PRR12
Protein A fragment: 1697-1711
Protein A site: PxxP_7
Protein A sequence: PAPPPKPETPEKTTS
Protein A construct: Bio Ado Ado-PAPPPKPETPEKTTS-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 9
Protein A protein: PRR12
Protein A fragment: 1720-1734
Protein A site: PxxP_8
Protein A sequence: ETAMPEPPAPEKPSL
Protein A construct: Bio Ado Ado-ETAMPEPPAPEKPSL-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: PRR12
Protein A fragment: 1143-1157
Protein A site: PxxP_9
Protein A sequence: CPPNPGPDGPRRRGR
Protein A construct: Bio Ado Ado-CPPNPGPDGPRRRGR-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: 3.62
Experiment series 11
Protein A protein: PRR12
Protein A fragment: 1161-1175
Protein A site: PxxP_10
Protein A sequence: KAKRDGPPRPRGRPR
Protein A construct: Bio Ado Ado-KAKRDGPPRPRGRPR-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: 3.52
Experiment series 12
Protein A protein: PRR12
Protein A fragment: 1309-1323
Protein A site: PxxP_11
Protein A sequence: PPKSVPPSVPARGLQ
Protein A construct: Bio Ado Ado-PPKSVPPSVPARGLQ-CONH2
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 3.72
Experiment series 13
Protein A protein: PRR12
Protein A fragment: 5-19
Protein A site: PxxP_12
Protein A modification: BAA86519.1
Protein A sequence: PGVPPGPPHPVRPPP
Protein A construct: Bio Ado Ado-PGVPPGPPHPVRPPP-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
Measured pKd: 3.48
Experiment series 14
Protein A protein: PRR12
Protein A fragment: 986-1000
Protein A site: PxxP_13
Protein A sequence: AYDPYGPYCPGRASG
Protein A construct: Bio Ado Ado-AYDPYGPYCPGRASG-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.