Details of experiments between the fragments P31947~1-248 and O43524~248-257
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: FOXO3
Protein B fragment: 248-257
Protein B site: 14-3-3_pSer253
Protein B modification: (SEP)253
Protein B sequence: RRRAVSMDNS
Protein B construct: RRRAV(pSer)MDNS (crude synthetic peptide)
Average holdup BI: -0.1
Holdup BI standard deviation: 1.39
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): 0.05
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.
Experiment series 2
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: FOXO3
Protein B fragment: 248-257
Protein B site: 14-3-3_pSer253
Protein B modification: (SEP)253
Protein B sequence: RRRAVSMDNS
Protein B construct: RRRAV(pSer)MDNS (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.7
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0
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.