Details of experiments between Q14155 and P30291
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: WEE1
Protein B fragment: 1-646
Protein B construct: endogenous protein (Jurkat)

Average holdup BI: 0.58
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): 2.65
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: 5.14

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: WEE1
Protein B fragment: 3-17
Protein B site: PxxP_1
Protein B sequence: FLSRQQPPPPRRAGA
Protein B construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2

Average holdup BI: 0.05
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: 2023/01/17
Number of measurements: 2
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: WEE1
Protein B fragment: 55-69
Protein B site: PxxP_2
Protein B sequence: FQEPDSPLPPARSPT
Protein B construct: Bio Ado Ado-FQEPDSPLPPARSPT-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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.