Details of experiments between Q8NI35 and P11169
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: Inadl
Protein A fragment: 1425-1530
Protein A site: PDZ_8
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A3
Protein B fragment: 487-496
Protein B site: PBM
Protein B sequence: EPAKETTTNV
Protein B construct: biotin-ado-ado-EPAKETTTNV
Average holdup BI: 0.06
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.59
Experiment series 2
Protein A protein: Inadl
Protein A fragment: 113-229
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A3
Protein B fragment: 487-496
Protein B site: PBM
Protein B sequence: EPAKETTTNV
Protein B construct: biotin-ado-ado-EPAKETTTNV
Average holdup BI: -0.01
Normalized holdup BI: -0.01
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 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: Inadl
Protein A fragment: 1673-1763
Protein A site: PDZ_10
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A3
Protein B fragment: 487-496
Protein B site: PBM
Protein B sequence: EPAKETTTNV
Protein B construct: biotin-ado-ado-EPAKETTTNV
Average holdup BI: 0.02
Normalized holdup BI: 0.02
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 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 4
Protein A protein: Inadl
Protein A fragment: 238-335
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A3
Protein B fragment: 487-496
Protein B site: PBM
Protein B sequence: EPAKETTTNV
Protein B construct: biotin-ado-ado-EPAKETTTNV
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: Crude peptide synthesis product. 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 5
Protein A protein: Inadl
Protein A fragment: 359-469
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A3
Protein B fragment: 487-496
Protein B site: PBM
Protein B sequence: EPAKETTTNV
Protein B construct: biotin-ado-ado-EPAKETTTNV
Average holdup BI: 0.02
Normalized holdup BI: 0.02
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 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 6
Protein A protein: Inadl
Protein A fragment: 547-651
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A3
Protein B fragment: 487-496
Protein B site: PBM
Protein B sequence: EPAKETTTNV
Protein B construct: biotin-ado-ado-EPAKETTTNV
Average holdup BI: -0.06
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 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 7
Protein A protein: Inadl
Protein A fragment: 673-780
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A3
Protein B fragment: 487-496
Protein B site: PBM
Protein B sequence: EPAKETTTNV
Protein B construct: biotin-ado-ado-EPAKETTTNV
Average holdup BI: 0.03
Normalized holdup BI: 0.03
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 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 8
Protein A protein: Inadl
Protein A fragment: 1058-1167
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A3
Protein B fragment: 487-496
Protein B site: PBM
Protein B sequence: EPAKETTTNV
Protein B construct: biotin-ado-ado-EPAKETTTNV
Average holdup BI: 0.02
Normalized holdup BI: 0.02
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 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 9
Protein A protein: Inadl
Protein A fragment: 1530-1615
Protein A site: PDZ_9
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A3
Protein B fragment: 487-496
Protein B site: PBM
Protein B sequence: EPAKETTTNV
Protein B construct: biotin-ado-ado-EPAKETTTNV
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: Crude peptide synthesis product. 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.