Details of experiments between P63104 and Q7L8J4
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
Protein B protein: SH3BP5L
Protein B fragment: 338-347
Protein B site: 14-3-3_pSer343
Protein B modification: (SEP)343
Protein B sequence: LRTVASDLQK
Protein B construct: LRTVA(pSer)DLQK (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.92
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: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
Protein B protein: SH3BP5L
Protein B fragment: 228-237
Protein B site: 14-3-3_pSer233
Protein B modification: (SEP)233
Protein B sequence: LKAQFSQILE
Protein B construct: LKAQF(pSer)QILE (crude synthetic peptide)
Average holdup BI: -0.32
Holdup BI standard deviation: 0.36
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.38
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 3
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
Protein B protein: SH3BP5L
Protein B fragment: 357-366
Protein B site: 14-3-3_pSer362
Protein B modification: (SEP)362
Protein B sequence: LSDHVSLDGQ
Protein B construct: LSDHV(pSer)LDGQ (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.59
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.