Details of experiments between Q14160 and P23634
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SCRIB
Protein A fragment: 1-1630
Protein A construct: endogenous protein (Jurkat)
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Experiment method: nHU-WB_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.08.03.
PUBMED ID of publication: 36542723
Number of measurements: 3
Measured pKd: 5.91
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Measured dissociation constant (10-6M): 9.44
Standard deviation of dissociation constant (10-6M): 0.59
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.03
Experiment series 3
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.43
Immobilized partner concentration in holdup experiment (10-6M): 7
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: 5.15
Experiment series 4
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.32
Immobilized partner concentration in holdup experiment (10-6M): 7
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: 4.92
Experiment series 5
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 7
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: 4.47
Experiment series 6
Protein A protein: SCRIB
Protein A fragment: 1090-1197
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 7
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: 4.38
Experiment series 7
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.6
Normalized holdup BI: 0.67
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: 5.11
Experiment series 8
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.59
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.62
Normalized holdup BI standard deviation: 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
PUBMED ID of publication: 36115835
Number of measurements: 4
Measured pKd: 5.03
Experiment series 9
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.45
Normalized holdup BI: 0.56
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: 4.9
Experiment series 10
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.56
Normalized holdup BI: 0.66
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 #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.1
Experiment series 11
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.66
Normalized holdup BI: 0.66
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 #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.1
Experiment series 12
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1232-1241
Protein B site: PBM
Protein B sequence: SSLQSLETSV
Protein B construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.55
Normalized holdup BI: 0.55
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 #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.89
Experiment series 13
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1-1241
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.23
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): 2.19
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: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B4
Protein B fragment: 1-1241
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.08
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): 1.06
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.