Details of experiments between O14924 and Q12959
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-Ahx-KTSAHHATFV
Protein B protein: DLG1
Protein B fragment: 314-412
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.16
Normalized holdup BI: 0.2
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: 1
Measured pKd: 4.15
Experiment series 2
Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-Ahx-KTSAHHATFV
Protein B protein: DLG1
Protein B fragment: 221-314
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.1
Normalized holdup BI: 0.13
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: 1
Measured pKd: 3.93
Experiment series 3
Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-Ahx-KTSAHHATFV
Protein B protein: DLG1
Protein B fragment: 457-570
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-ado-ado-KTSAHHATFV
Protein B protein: DLG1
Protein B fragment: 221-314
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.14
Normalized holdup BI: 0.17
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: 4.07
Experiment series 5
Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-ado-ado-KTSAHHATFV
Protein B protein: DLG1
Protein B fragment: 314-412
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.28
Normalized holdup BI: 0.31
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: 4.43
Experiment series 6
Protein A protein: RGS12
Protein A fragment: 1438-1447
Protein A site: PBM
Protein A sequence: KTSAHHATFV
Protein A construct: Biotin-ado-ado-KTSAHHATFV
Protein B protein: DLG1
Protein B fragment: 457-570
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.04
Normalized holdup BI: 0.06
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: 3.56