Details of experiments between Q14160 and P11166
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: SLC2A1
Protein B fragment: 483-492
Protein B site: PBM
Protein B sequence: FHPLGADSQV
Protein B construct: biotin-ttds-FHPLGADSQV
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: 3.95
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: SLC2A1
Protein B fragment: 483-492
Protein B site: PBM
Protein B sequence: FHPLGADSQV
Protein B construct: biotin-ttds-FHPLGADSQV
Average holdup BI: 0.15
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.16
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: 2
Measured pKd: 4.02
Experiment series 3
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A1
Protein B fragment: 483-492
Protein B site: PBM
Protein B sequence: FHPLGADSQV
Protein B construct: biotin-ttds-FHPLGADSQV
Average holdup BI: 0.06
Holdup BI standard deviation: 0.06
Normalized holdup BI: 0.06
Normalized holdup BI standard deviation: 0.07
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: 3.58
Experiment series 4
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLC2A1
Protein B fragment: 483-492
Protein B site: PBM
Protein B sequence: FHPLGADSQV
Protein B construct: biotin-ttds-FHPLGADSQV
Average holdup BI: 0.01
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.01
Normalized holdup BI standard deviation: 0.05
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
The affinity was below the detection threshold of the assay.
Experiment series 5
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: SLC2A1
Protein B fragment: 1-492
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: -0
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): 0.04
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 6
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: SLC2A1
Protein B fragment: 1-492
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: -0.02
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): 0.13
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.