Details of experiments between the fragments P61981~1-247 and Q9Y2U9~205-214
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KLHDC2
Protein B fragment: 205-214
Protein B site: 14-3-3_pThr210
Protein B modification: (TPO)210
Protein B sequence: WSQPITTGKA
Protein B construct: WSQPI(pThr)TGKA (crude synthetic peptide)
Average holdup BI: 0.26
Holdup BI standard deviation: 0.06
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): 3.87
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
Measured pKd: 3.51
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: KLHDC2
Protein B fragment: 205-214
Protein B site: 14-3-3_pThr210
Protein B modification: (TPO)210
Protein B sequence: WSQPITTGKA
Protein B construct: WSQPI(pThr)TGKA (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.06
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): 3.78
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 construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KLHDC2
Protein B fragment: 205-214
Protein B site: 14-3-3_pThr210
Protein B modification: (TPO)210
Protein B sequence: WSQPITTGKA
Protein B construct: WSQPI(pThr)TGKA (crude synthetic peptide)
Average holdup BI: 0.22
Holdup BI standard deviation: 0.05
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): 3.63
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.