Details of experiments between Q8TEW0 and Q14155
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: PARD3
Protein A fragment: 1-1356
Protein A construct: endogenous protein (Jurkat)

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

Average holdup BI: -0.01
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): 0.07
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: PARD3
Protein A fragment: 582-685
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: ARHGEF7
Protein B fragment: 794-803
Protein B site: PBM
Protein B sequence: NDPAWDETNL
Protein B construct: biotin-ttds-NDPAWDETNL

Average holdup BI: 0.19
Holdup BI standard deviation: 0.06
Normalized holdup BI: 0.19
Normalized holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 17
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 4.16