Details of experiments between Q04917 and Q9UQL6
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: HDAC5
Protein B fragment: 254-263
Protein B site: 14-3-3_pSer259
Protein B modification: (SEP)259
Protein B sequence: LRKTASEPNL
Protein B construct: LRKTA(pSer)EPNL (crude synthetic peptide)
Average holdup BI: 0.93
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.95
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
Measured pKd: 4.99
Experiment series 2
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: HDAC5
Protein B fragment: 542-551
Protein B site: 14-3-3_pThr547
Protein B modification: (TPO)547
Protein B sequence: PRQPTTHPEE
Protein B construct: PRQPT(pThr)HPEE (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.94
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.