Details of experiments between Q14160 and P35222
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: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CTNNB1
Protein B fragment: 772-781
Protein B site: PBM
Protein B sequence: NQLAWFDTDL
Protein B construct: biotin-ttds-NQLAWFDTDL
Average holdup BI: 0.51
Normalized holdup BI: 0.53
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: 4.86
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CTNNB1
Protein B fragment: 772-781
Protein B site: PBM
Protein B sequence: NQLAWFDTDL
Protein B construct: biotin-ttds-NQLAWFDTDL
Average holdup BI: 0.46
Normalized holdup BI: 0.52
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: 4.82
Experiment series 3
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: CTNNB1
Protein B fragment: 772-781
Protein B site: PBM
Protein B sequence: NQLAWFDTDL
Protein B construct: biotin-ttds-NQLAWFDTDL
Average holdup BI: 0.05
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.06
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: 3.57
Experiment series 4
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: CTNNB1
Protein B fragment: 1-781
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.21
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): 0.82
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 5
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: CTNNB1
Protein B fragment: 1-781
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: -0.03
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): 0.11
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.