Details of experiments between Q96DX4 and O00499
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: RSPRY1
Protein A fragment: 1-576
Protein A construct: endogenous protein (Jurkat)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.66
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): 3.79
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: 5.29
Experiment series 2
Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.31
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: 4.43
Experiment series 3
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.79
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: 5.41
Experiment series 4
Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.13
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: 3.93
Experiment series 5
Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.33
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: 4.46
Experiment series 6
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.8
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: 5.43
Experiment series 7
Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.15
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: 3.99
Experiment series 8
Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.38
Holdup BI standard deviation: 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: 2023/01/17
Number of measurements: 2
Measured pKd: 4.56
Experiment series 9
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.75
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 5.29
Experiment series 10
Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 4.15
Experiment series 11
Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.07
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: 2023/01/17
Number of measurements: 2
Measured pKd: 3.6
Experiment series 12
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.46
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 4.73
Experiment series 13
Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 14
Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.09
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 13
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 2
Measured pKd: 3.88
Experiment series 15
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.66
Immobilized partner concentration in holdup experiment (10-6M): 13
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 5.27
Experiment series 16
Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 13
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 17
Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.14
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 8
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 2
Measured pKd: 4.34
Experiment series 18
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.6
Immobilized partner concentration in holdup experiment (10-6M): 8
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 5.36
Experiment series 19
Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 8
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 20
Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.01
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: 2023/01/17
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 21
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 22
Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 23
Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.22
Holdup BI standard deviation: 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: 2
Measured pKd: 4.22
Experiment series 24
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.71
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: 5.22
Experiment series 25
Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
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 26
Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.11
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 2
Measured pKd: 4.12
Experiment series 27
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.62
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 5.34
Experiment series 28
Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
Measured pKd: 3.89
Experiment series 29
Protein A protein: RSPRY1
Protein A fragment: 71-85
Protein A site: PxxP_1
Protein A sequence: ADTRSQPRDPVRPPR
Protein A construct: Bio Ado Ado-ADTRSQPRDPVRPPR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.33
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
Measured pKd: 4.46
Experiment series 30
Protein A protein: RSPRY1
Protein A fragment: 74-88
Protein A site: PxxP_2
Protein A sequence: RSQPRDPVRPPRRGR
Protein A construct: Bio Ado Ado-RSQPRDPVRPPRRGR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.75
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: 5.29
Experiment series 31
Protein A protein: RSPRY1
Protein A fragment: 84-98
Protein A site: PxxP_3
Protein A sequence: PRRGRGPHEPRRKKQ
Protein A construct: Bio Ado Ado-PRRGRGPHEPRRKKQ-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.15
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: 4.02