Details of experiments between P61981 and P04049
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: RAF1
Protein B fragment: 254-264
Protein B site: 14-3-3_pSer259
Protein B modification: (SEP)259
Protein B sequence: RQRSTSTPNV
Protein B construct: RQRST(pSer)TPNVH (crude synthetic peptide)

Average holdup BI: 0.24
Holdup BI standard deviation: 0.65
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): 0.27
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: RAF1
Protein B fragment: 254-264
Protein B site: 14-3-3_pSer259
Protein B modification: (SEP)259
Protein B sequence: RQRSTSTPNV
Protein B construct: RQRST(pSer)TPNVH (crude synthetic peptide)

Average holdup BI: 0.04
Holdup BI standard deviation: 0.4
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): 0.07
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.