Details of experiments between the fragments P25054~2834-2843 and Q6P0Q8~1102-1193
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: APC
Protein A fragment: 2834-2843
Protein A site: PBM
Protein A sequence: HSGSYLVTSV
Protein A construct: Biotin-Ahx-HSGSYLVTSV
Protein B protein: MAST2
Protein B fragment: 1102-1193
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.66
Holdup BI standard deviation: 0
Normalized holdup BI: 0.66
Normalized holdup BI standard deviation: 0
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: The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 5.11
Experiment series 2
Protein A protein: APC
Protein A fragment: 2834-2843
Protein A site: PBM
Protein A sequence: HSGSYLVTSV
Protein A construct: Biotin-Ahx-HSGSYLVTSV
Protein B protein: MAST2
Protein B fragment: 1102-1193
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.74
Normalized holdup BI: 0.74
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.27