Details of experiments between Q8N448 and Q7Z6J2
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: LNX2
Protein A fragment: 681-690
Protein A site: PBM
Protein A sequence: TVICWPGSLV
Protein A construct: Biotin-Ahx-TVICWPGSLV
Protein B protein: GRASP
Protein B fragment: 96-193
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.39
Normalized holdup BI: 0.39
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: 4.59
Experiment series 2
Protein A protein: LNX2
Protein A fragment: 334-426
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: GRASP
Protein B fragment: 386-395
Protein B site: PBM
Protein B sequence: RSLEEEESQL
Protein B construct: biotin-ttds-RSLEEEESQL
Average holdup BI: -0.04
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: LNX2
Protein A fragment: 459-560
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: GRASP
Protein B fragment: 386-395
Protein B site: PBM
Protein B sequence: RSLEEEESQL
Protein B construct: biotin-ttds-RSLEEEESQL
Average holdup BI: 0.03
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: LNX2
Protein A fragment: 592-688
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: GRASP
Protein B fragment: 386-395
Protein B site: PBM
Protein B sequence: RSLEEEESQL
Protein B construct: biotin-ttds-RSLEEEESQL
Average holdup BI: 0.06
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 5
Protein A protein: LNX2
Protein A fragment: 334-426
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: GRASP
Protein B fragment: 386-395
Protein B site: PBM
Protein B sequence: RSLEEEESQL
Protein B construct: biotin-ttds-RSLEEEESQL
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
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: LNX2
Protein A fragment: 459-560
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: GRASP
Protein B fragment: 386-395
Protein B site: PBM
Protein B sequence: RSLEEEESQL
Protein B construct: biotin-ttds-RSLEEEESQL
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.