Details of experiments between Q8TBC3 and Q5T2W1
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SHKBP1
Protein A fragment: 698-707
Protein A site: PBM
Protein A sequence: PKMKLNETSF
Protein A construct: Biotin-Ahx-PKMKLNETSF
Protein B protein: NHERF3
Protein B fragment: 2-107
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.34
Normalized holdup BI: 0.37
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: 1
Measured pKd: 4.55
Experiment series 2
Protein A protein: SHKBP1
Protein A fragment: 698-707
Protein A site: PBM
Protein A sequence: PKMKLNETSF
Protein A construct: Biotin-Ahx-PKMKLNETSF
Protein B protein: NHERF3
Protein B fragment: 238-329
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0
Normalized holdup BI: -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: 1
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: SHKBP1
Protein A fragment: 698-707
Protein A site: PBM
Protein A sequence: PKMKLNETSF
Protein A construct: Biotin-Ahx-PKMKLNETSF
Protein B protein: NHERF3
Protein B fragment: 375-459
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.04
Normalized holdup BI: 0.04
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: 1
The affinity was below the detection threshold of the assay.