Details of experiments between Q96L92 and Q96KB5
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PBK
Protein B fragment: 313-322
Protein B site: PBM
Protein B sequence: HIVEALETDV
Protein B construct: biotin-ttds-HIVEALETDV
Average holdup BI: 0.19
Immobilized partner concentration in holdup experiment (10-6M): 5.5
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: 4.7
Experiment series 2
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PBK
Protein B fragment: 313-322
Protein B site: PBM
Protein B sequence: HIVEALETDV
Protein B construct: biotin-ttds-HIVEALETDV
Average holdup BI: 0.4
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.4
Normalized holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 11.6
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: 4.83
Experiment series 3
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: PBK
Protein B fragment: 1-322
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): 0.66
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.27.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 6))
PUBMED ID of publication: 36542723
Number of measurements: 6
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: SNX27
Protein A fragment: 1-541
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SNX27
Protein B protein: PBK
Protein B fragment: 1-322
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): 0.87
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.27.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 6))
PUBMED ID of publication: 36542723
Number of measurements: 6
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: PBK
Protein B fragment: 1-322
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): 0.98
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.