Details of experiments between Q14160 and Q13574
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: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Experiment method: nHU-WB_Jurkat
Details of affinity fitting: Hill equation (8 point titration) h=0.6
Date of measurement: 2022.08.03.
PUBMED ID of publication: 36542723
Number of measurements: 3
Measured pKd: 6.4
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 1-1630
Protein A construct: endogenous protein (Jurkat)
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
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: 6.42
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: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Measured dissociation constant (10-6M): 36.87
Standard deviation of dissociation constant (10-6M): 3.4
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.43
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: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.25
Immobilized partner concentration in holdup experiment (10-6M): 6
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: 4.81
Experiment series 5
Protein A protein: SCRIB
Protein A fragment: 1090-1197
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 6
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: 4.4
Experiment series 6
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 6
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: 4.36
Experiment series 7
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 6
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 8
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.5
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.56
Normalized holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 13.5
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: 5.05
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: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.37
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.46
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 13.5
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: 3
Measured pKd: 4.86
Experiment series 10
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.36
Holdup BI standard deviation: 0.24
Normalized holdup BI: 0.38
Normalized holdup BI standard deviation: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 13.5
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: 5
Measured pKd: 4.71
Experiment series 11
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.47
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 12
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.32
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 #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.45
Experiment series 13
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.45
Normalized holdup BI: 0.45
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: 4.71
Experiment series 14
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: DGKZ
Protein B fragment: 1-928
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: -0.09
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): 0.43
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 15
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: DGKZ
Protein B fragment: 1-928
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: -0.06
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): 0.27
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.