Details of experiments between the fragments P61981~1-247 and Q08AE8~459-468
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: SPIRE1
Protein B fragment: 459-468
Protein B site: 14-3-3_pSer464
Protein B modification: (SEP)464
Protein B sequence: LAELDSSESE
Protein B construct: LAELD(pSer)SESE (crude synthetic peptide)
Average holdup BI: -0.32
Holdup BI standard deviation: 0.86
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.4
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: SPIRE1
Protein B fragment: 459-468
Protein B site: 14-3-3_pSer464
Protein B modification: (SEP)464
Protein B sequence: LAELDSSESE
Protein B construct: LAELD(pSer)SESE (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.39
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.1
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.