Details of experiments between P62258 and Q9BZF3
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: OSBPL6
Protein B fragment: 285-294
Protein B site: 14-3-3_pSer290
Protein B modification: (SEP)290
Protein B sequence: LQRTQSAPNF
Protein B construct: LQRTQ(pSer)APNF (crude synthetic peptide)
Average holdup BI: 0.83
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.61
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
Measured pKd: 4.55
Experiment series 2
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: OSBPL6
Protein B fragment: 285-294
Protein B site: 14-3-3_pSer290
Protein B modification: (SEP)290
Protein B sequence: LQRTQSAPNF
Protein B construct: LQRTQ(pSer)APNF (crude synthetic peptide)
Average holdup BI: 0.81
Holdup BI standard deviation: 0.32
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.44
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: 4.56
Experiment series 3
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: OSBPL6
Protein B fragment: 39-48
Protein B site: 14-3-3_pSer44
Protein B modification: (SEP)44
Protein B sequence: LERTASSSTE
Protein B construct: LERTA(pSer)SSTE (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.23
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.73
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 4
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: OSBPL6
Protein B fragment: 39-48
Protein B site: 14-3-3_pSer44
Protein B modification: (SEP)44
Protein B sequence: LERTASSSTE
Protein B construct: LERTA(pSer)SSTE (crude synthetic peptide)
Average holdup BI: -0.3
Holdup BI standard deviation: 0.49
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.69
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.