Details of experiments between O14907 and P20020
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: TX1BP3
Protein A fragment: 7-124
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: biotin-ttds-SPLHSLETSL
Average holdup BI: 0.29
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.32
Normalized holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
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: 4.45
Experiment series 2
Protein A protein: TX1BP3
Protein A fragment: 7-124
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: Biotin-ado-ado-SPLHSLETSL
Average holdup BI: 0.5
Normalized holdup BI: 0.5
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.79
Experiment series 3
Protein A protein: TX1BP3
Protein A fragment: 7-124
Protein A site: PDZ
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1-1220
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.34
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): 1.55
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.08.03.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
PUBMED ID of publication: 36542723
Number of measurements: 3
The affinity was below the detection threshold of the assay.