Details of experiments between P62258 and Q15032
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: R3HDM1
Protein B fragment: 376-385
Protein B site: 14-3-3_pSer381
Protein B modification: (SEP)381
Protein B sequence: VTKASSFSGI
Protein B construct: VTKAS(pSer)FSGI (crude synthetic peptide)
Average holdup BI: 0.53
Holdup BI standard deviation: 0.04
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): 6.17
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: 3.89
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: R3HDM1
Protein B fragment: 376-385
Protein B site: 14-3-3_pSer381
Protein B modification: (SEP)381
Protein B sequence: VTKASSFSGI
Protein B construct: VTKAS(pSer)FSGI (crude synthetic peptide)
Average holdup BI: 0.57
Holdup BI standard deviation: 0.06
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): 5.32
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: 4.07
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: R3HDM1
Protein B fragment: 1093-1102
Protein B site: 14-3-3_pSer1098
Protein B modification: (SEP)1098
Protein B sequence: LERASSQ---
Protein B construct: FHILERAS(pSer)Q (crude synthetic peptide)
Average holdup BI: 0.32
Holdup BI standard deviation: 0.06
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): 4.73
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: 3.51
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: R3HDM1
Protein B fragment: 1093-1102
Protein B site: 14-3-3_pSer1098
Protein B modification: (SEP)1098
Protein B sequence: LERASSQ---
Protein B construct: FHILERAS(pSer)Q (crude synthetic peptide)
Average holdup BI: 0.27
Holdup BI standard deviation: 0.05
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): 4.09
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 5
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: R3HDM1
Protein B fragment: 294-303
Protein B site: 14-3-3_pSer299
Protein B modification: (SEP)299
Protein B sequence: RDRIFSQDSL
Protein B construct: RDRIF(pSer)QDSL (crude synthetic peptide)
Average holdup BI: -0.39
Holdup BI standard deviation: 0.28
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): 2.01
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 6
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: R3HDM1
Protein B fragment: 1092-1101
Protein B site: 14-3-3_pSer1097
Protein B modification: (SEP)1097
Protein B sequence: ILERASSQ--
Protein B construct: FHILERA(pSer)SQ (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.13
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): 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
Measured pKd: 3.15
Experiment series 7
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: R3HDM1
Protein B fragment: 350-359
Protein B site: 14-3-3_pSer355
Protein B modification: (SEP)355
Protein B sequence: NELKYSEPRP
Protein B construct: NELKY(pSer)EPRP (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.09
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): 0.66
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 8
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: R3HDM1
Protein B fragment: 275-284
Protein B site: 14-3-3_pSer280
Protein B modification: (SEP)280
Protein B sequence: KDDRRSKSIE
Protein B construct: KDDRR(pSer)KSIE (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.09
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): 0.58
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 9
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: R3HDM1
Protein B fragment: 294-303
Protein B site: 14-3-3_pSer299
Protein B modification: (SEP)299
Protein B sequence: RDRIFSQDSL
Protein B construct: RDRIF(pSer)QDSL (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.14
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): 0.49
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 10
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: R3HDM1
Protein B fragment: 1092-1101
Protein B site: 14-3-3_pSer1097
Protein B modification: (SEP)1097
Protein B sequence: ILERASSQ--
Protein B construct: FHILERA(pSer)SQ (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.13
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): 0.44
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
The affinity was below the detection threshold of the assay.
Experiment series 11
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: R3HDM1
Protein B fragment: 137-146
Protein B site: 14-3-3_pSer142
Protein B modification: (SEP)142
Protein B sequence: LSRDSSQEYT
Protein B construct: LSRDS(pSer)QEYT (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.05
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): 0.26
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 12
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: R3HDM1
Protein B fragment: 275-284
Protein B site: 14-3-3_pSer280
Protein B modification: (SEP)280
Protein B sequence: KDDRRSKSIE
Protein B construct: KDDRR(pSer)KSIE (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.17
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): 0.17
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 13
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: R3HDM1
Protein B fragment: 350-359
Protein B site: 14-3-3_pSer355
Protein B modification: (SEP)355
Protein B sequence: NELKYSEPRP
Protein B construct: NELKY(pSer)EPRP (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.1
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): 0.16
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 14
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: R3HDM1
Protein B fragment: 137-146
Protein B site: 14-3-3_pSer142
Protein B modification: (SEP)142
Protein B sequence: LSRDSSQEYT
Protein B construct: LSRDS(pSer)QEYT (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.06
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): 0.04
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.