Details of experiments between Q12959 and Q13224
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: GRIN2B
Protein B fragment: 1475-1484
Protein B site: PBM
Protein B sequence: EKLSSIESDV
Protein B construct: Biotin-Ahx-EKLSSIESDV
Average holdup BI: 0.5
Normalized holdup BI: 0.59
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: 4.97
Experiment series 2
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: GRIN2B
Protein B fragment: 1475-1484
Protein B site: PBM
Protein B sequence: EKLSSIESDV
Protein B construct: Biotin-Ahx-EKLSSIESDV
Average holdup BI: 0.2
Normalized holdup BI: 0.25
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: 4.3
Experiment series 3
Protein A protein: DLG1
Protein A fragment: 457-570
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: GRIN2B
Protein B fragment: 1475-1484
Protein B site: PBM
Protein B sequence: EKLSSIESDV
Protein B construct: Biotin-Ahx-EKLSSIESDV
Average holdup BI: 0.03
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.