Details of experiments between Q14155 and Q6F5E8
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3

Protein B protein: CARMIL2
Protein B fragment: 1-1435
Protein B construct: endogenous protein (Jurkat)

Average holdup BI: 0.22
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): 4.84
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
The affinity was below the detection threshold of the assay.


Experiment series 2

Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: CARMIL2
Protein B fragment: 1030-1044
Protein B site: PxxP_1
Protein B sequence: HHHRPPPGGPQVPPA
Protein B construct: Bio Ado Ado-HHHRPPPGGPQVPPA-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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 3

Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: CARMIL2
Protein B fragment: 1133-1147
Protein B site: PxxP_2
Protein B sequence: GDLLRPPTRPSRGEE
Protein B construct: Bio Ado Ado-GDLLRPPTRPSRGEE-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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 4

Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3

Protein B protein: CARMIL2
Protein B fragment: 1408-1422
Protein B site: PxxP_3
Protein B sequence: PPQPTEPSSPERSPP
Protein B construct: Bio Ado Ado-PPQPTEPSSPERSPP-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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.