Details of experiments between the fragments P61981~1-247 and Q15772~1167-1176
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: SPEG
Protein B fragment: 1167-1176
Protein B site: 14-3-3_pSer1172
Protein B modification: (SEP)1172
Protein B sequence: LEKMPSIPEE
Protein B construct: LEKMP(pSer)IPEE (crude synthetic peptide)
Average holdup BI: -0.16
Holdup BI standard deviation: 0.27
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): 0.58
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 construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: SPEG
Protein B fragment: 1167-1176
Protein B site: 14-3-3_pSer1172
Protein B modification: (SEP)1172
Protein B sequence: LEKMPSIPEE
Protein B construct: LEKMP(pSer)IPEE (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.14
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: 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: SPEG
Protein B fragment: 1167-1176
Protein B site: 14-3-3_pSer1172
Protein B modification: (SEP)1172
Protein B sequence: LEKMPSIPEE
Protein B construct: LEKMP(pSer)IPEE (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.28
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.