Details of experiments between Q12959 and O15439
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: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
Average holdup BI: 0.21
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: 4.48
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: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
Average holdup BI: 0.2
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: 4.45
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: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
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: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
Average holdup BI: 0.34
Holdup BI standard deviation: 0.1
Normalized holdup BI: 0.38
Normalized holdup BI standard deviation: 0.11
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: 2
Measured pKd: 4.57
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: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
Average holdup BI: 0.27
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0.32
Normalized holdup BI standard deviation: 0.11
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: 2
Measured pKd: 4.46
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: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
Average holdup BI: 0.24
Holdup BI standard deviation: 0.1
Normalized holdup BI: 0.29
Normalized holdup BI standard deviation: 0.12
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: 2
Measured pKd: 4.38
Experiment series 7
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
Average holdup BI: 0.22
Normalized holdup BI: 0.29
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: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 4.38
Experiment series 8
Protein A protein: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
Average holdup BI: 0.27
Normalized holdup BI: 0.32
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: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 4.45
Experiment series 9
Protein A protein: DLG1
Protein A fragment: 457-570
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: biotin-ttds-STLTIFETAL
Average holdup BI: 0.18
Normalized holdup BI: 0.24
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: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 4.26
Experiment series 10
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: Biotin-ado-ado-STLTIFETAL
Average holdup BI: 0.26
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: 4.42
Experiment series 11
Protein A protein: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: Biotin-ado-ado-STLTIFETAL
Average holdup BI: 0.39
Normalized holdup BI: 0.44
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: 4.68
Experiment series 12
Protein A protein: DLG1
Protein A fragment: 457-570
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1316-1325
Protein B site: PBM
Protein B sequence: STLTIFETAL
Protein B construct: Biotin-ado-ado-STLTIFETAL
Average holdup BI: 0.18
Normalized holdup BI: 0.27
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: 4.34
Experiment series 13
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1-1325
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: -0.15
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.92
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.08.03.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
PUBMED ID of publication: 36542723
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 14
Protein A protein: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: ABCC4
Protein B fragment: 1-1325
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: -0.05
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.12
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.08.03.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
PUBMED ID of publication: 36542723
Number of measurements: 3
The affinity was below the detection threshold of the assay.