Details of experiments between Q6F5E8 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: CARMIL2
Protein A fragment: 1-1435
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: 0.32
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): 6.16
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: 4.66


Experiment series 2

Protein A protein: CARMIL2
Protein A fragment: 1030-1044
Protein A site: PxxP_1
Protein A sequence: HHHRPPPGGPQVPPA
Protein A construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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
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 3

Protein A protein: CARMIL2
Protein A fragment: 1133-1147
Protein A site: PxxP_2
Protein A sequence: GDLLRPPTRPSRGEE
Protein A construct: Bio Ado Ado-GDLLRPPTRPSRGEE-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 3.6


Experiment series 4

Protein A protein: CARMIL2
Protein A fragment: 1408-1422
Protein A site: PxxP_3
Protein A sequence: PPQPTEPSSPERSPP
Protein A construct: Bio Ado Ado-PPQPTEPSSPERSPP-CONH2

Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.02
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.


Experiment series 5

Protein A protein: CARMIL2
Protein A fragment: 1030-1044
Protein A site: PxxP_1
Protein A sequence: HHHRPPPGGPQVPPA
Protein A construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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
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 6

Protein A protein: CARMIL2
Protein A fragment: 1133-1147
Protein A site: PxxP_2
Protein A sequence: GDLLRPPTRPSRGEE
Protein A construct: Bio Ado Ado-GDLLRPPTRPSRGEE-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: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 3.5


Experiment series 7

Protein A protein: CARMIL2
Protein A fragment: 1408-1422
Protein A site: PxxP_3
Protein A sequence: PPQPTEPSSPERSPP
Protein A construct: Bio Ado Ado-PPQPTEPSSPERSPP-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: -0.03
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 8

Protein A protein: CARMIL2
Protein A fragment: 1030-1044
Protein A site: PxxP_1
Protein A sequence: HHHRPPPGGPQVPPA
Protein A construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 9

Protein A protein: CARMIL2
Protein A fragment: 1133-1147
Protein A site: PxxP_2
Protein A sequence: GDLLRPPTRPSRGEE
Protein A construct: Bio Ado Ado-GDLLRPPTRPSRGEE-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: 0.19
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: 4.13


Experiment series 10

Protein A protein: CARMIL2
Protein A fragment: 1408-1422
Protein A site: PxxP_3
Protein A sequence: PPQPTEPSSPERSPP
Protein A construct: Bio Ado Ado-PPQPTEPSSPERSPP-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: -0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 11

Protein A protein: CARMIL2
Protein A fragment: 1030-1044
Protein A site: PxxP_1
Protein A sequence: HHHRPPPGGPQVPPA
Protein A construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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: -0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 12

Protein A protein: CARMIL2
Protein A fragment: 1133-1147
Protein A site: PxxP_2
Protein A sequence: GDLLRPPTRPSRGEE
Protein A construct: Bio Ado Ado-GDLLRPPTRPSRGEE-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 13

Protein A protein: CARMIL2
Protein A fragment: 1408-1422
Protein A site: PxxP_3
Protein A sequence: PPQPTEPSSPERSPP
Protein A construct: Bio Ado Ado-PPQPTEPSSPERSPP-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: -0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 14

Protein A protein: CARMIL2
Protein A fragment: 1030-1044
Protein A site: PxxP_1
Protein A sequence: HHHRPPPGGPQVPPA
Protein A construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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
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 15

Protein A protein: CARMIL2
Protein A fragment: 1133-1147
Protein A site: PxxP_2
Protein A sequence: GDLLRPPTRPSRGEE
Protein A construct: Bio Ado Ado-GDLLRPPTRPSRGEE-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: -0.01
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 16

Protein A protein: CARMIL2
Protein A fragment: 1408-1422
Protein A site: PxxP_3
Protein A sequence: PPQPTEPSSPERSPP
Protein A construct: Bio Ado Ado-PPQPTEPSSPERSPP-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: CARMIL2
Protein A fragment: 1030-1044
Protein A site: PxxP_1
Protein A sequence: HHHRPPPGGPQVPPA
Protein A construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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: 0.01
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 18

Protein A protein: CARMIL2
Protein A fragment: 1133-1147
Protein A site: PxxP_2
Protein A sequence: GDLLRPPTRPSRGEE
Protein A construct: Bio Ado Ado-GDLLRPPTRPSRGEE-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: 0.02
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 19

Protein A protein: CARMIL2
Protein A fragment: 1408-1422
Protein A site: PxxP_3
Protein A sequence: PPQPTEPSSPERSPP
Protein A construct: Bio Ado Ado-PPQPTEPSSPERSPP-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
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: CARMIL2
Protein A fragment: 1030-1044
Protein A site: PxxP_1
Protein A sequence: HHHRPPPGGPQVPPA
Protein A construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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: 0.03
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: 1
The affinity was below the detection threshold of the assay.


Experiment series 21

Protein A protein: CARMIL2
Protein A fragment: 1133-1147
Protein A site: PxxP_2
Protein A sequence: GDLLRPPTRPSRGEE
Protein A construct: Bio Ado Ado-GDLLRPPTRPSRGEE-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: -0.1
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 22

Protein A protein: CARMIL2
Protein A fragment: 1408-1422
Protein A site: PxxP_3
Protein A sequence: PPQPTEPSSPERSPP
Protein A construct: Bio Ado Ado-PPQPTEPSSPERSPP-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: CARMIL2
Protein A fragment: 1030-1044
Protein A site: PxxP_1
Protein A sequence: HHHRPPPGGPQVPPA
Protein A construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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: 0.01
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 24

Protein A protein: CARMIL2
Protein A fragment: 1133-1147
Protein A site: PxxP_2
Protein A sequence: GDLLRPPTRPSRGEE
Protein A construct: Bio Ado Ado-GDLLRPPTRPSRGEE-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: -0.01
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 25

Protein A protein: CARMIL2
Protein A fragment: 1408-1422
Protein A site: PxxP_3
Protein A sequence: PPQPTEPSSPERSPP
Protein A construct: Bio Ado Ado-PPQPTEPSSPERSPP-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: CARMIL2
Protein A fragment: 1030-1044
Protein A site: PxxP_1
Protein A sequence: HHHRPPPGGPQVPPA
Protein A construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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: -0.02
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
The affinity was below the detection threshold of the assay.


Experiment series 27

Protein A protein: CARMIL2
Protein A fragment: 1133-1147
Protein A site: PxxP_2
Protein A sequence: GDLLRPPTRPSRGEE
Protein A construct: Bio Ado Ado-GDLLRPPTRPSRGEE-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
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
The affinity was below the detection threshold of the assay.


Experiment series 28

Protein A protein: CARMIL2
Protein A fragment: 1408-1422
Protein A site: PxxP_3
Protein A sequence: PPQPTEPSSPERSPP
Protein A construct: Bio Ado Ado-PPQPTEPSSPERSPP-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: -0.02
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
The affinity was below the detection threshold of the assay.


Experiment series 29

Protein A protein: CARMIL2
Protein A fragment: 1030-1044
Protein A site: PxxP_1
Protein A sequence: HHHRPPPGGPQVPPA
Protein A construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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: 0.04
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 30

Protein A protein: CARMIL2
Protein A fragment: 1133-1147
Protein A site: PxxP_2
Protein A sequence: GDLLRPPTRPSRGEE
Protein A construct: Bio Ado Ado-GDLLRPPTRPSRGEE-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: 0.03
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 31

Protein A protein: CARMIL2
Protein A fragment: 1408-1422
Protein A site: PxxP_3
Protein A sequence: PPQPTEPSSPERSPP
Protein A construct: Bio Ado Ado-PPQPTEPSSPERSPP-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: -0.02
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: 1
The affinity was below the detection threshold of the assay.