Details of experiments between Q12959 and A4U7F2
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: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.25
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.58
Experiment series 2
Protein A protein: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
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: DLG1
Protein A fragment: 457-570
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal 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: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.59
Normalized holdup BI: 0.66
Immobilized partner concentration in holdup experiment (10-6M): 22
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: 5.01
Experiment series 5
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.34
Normalized holdup BI: 0.4
Immobilized partner concentration in holdup experiment (10-6M): 22
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: 4.52
Experiment series 6
Protein A protein: DLG1
Protein A fragment: 457-570
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.19
Normalized holdup BI: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 22
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: 4.13