Details of experiments between the fragments P31947~1-248 and Q86SQ0~237-246
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: PHLDB2
Protein B fragment: 237-246
Protein B site: 14-3-3_pSer242
Protein B modification: (SEP)242
Protein B sequence: RTRKYSSSSL
Protein B construct: RTRKY(pSer)SSSL (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.56
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: PHLDB2
Protein B fragment: 237-246
Protein B site: 14-3-3_pSer242
Protein B modification: (SEP)242
Protein B sequence: RTRKYSSSSL
Protein B construct: RTRKY(pSer)SSSL (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.61
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.