Details of experiments between P62258 and P15056
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: BRAF
Protein B fragment: 724-733
Protein B site: 14-3-3_pSer729
Protein B modification: (SEP)729
Protein B sequence: IHRSASEPSL
Protein B construct: IHRSA(pSer)EPSL (crude synthetic peptide)
Average holdup BI: 0.79
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): 6.83
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: 4.43
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: BRAF
Protein B fragment: 465-474
Protein B site: 14-3-3_pThr470
Protein B modification: (TPO)470
Protein B sequence: SGSFGTVYKG
Protein B construct: SGSFG(pThr)VYKG (crude synthetic peptide)
Average holdup BI: 0.26
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): 4.99
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: 3.38
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: BRAF
Protein B fragment: 465-474
Protein B site: 14-3-3_pThr470
Protein B modification: (TPO)470
Protein B sequence: SGSFGTVYKG
Protein B construct: SGSFG(pThr)VYKG (crude synthetic peptide)
Average holdup BI: 0.2
Holdup BI standard deviation: 0.04
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): 4.21
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: BRAF
Protein B fragment: 334-343
Protein B site: 14-3-3_pSer339
Protein B modification: (SEP)339
Protein B sequence: PSPSKSIPIP
Protein B construct: PSPSK(pSer)IPIP (crude synthetic peptide)
Average holdup BI: 0.25
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): 4.16
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: 3.37
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: BRAF
Protein B fragment: 360-369
Protein B site: 14-3-3_pSer365
Protein B modification: (SEP)365
Protein B sequence: RDRSSSAPNV
Protein B construct: RDRSS(pSer)APNV (crude synthetic peptide)
Average holdup BI: 0.72
Holdup BI standard deviation: 0.12
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): 3.22
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: 4.35
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: BRAF
Protein B fragment: 462-471
Protein B site: 14-3-3_pSer467
Protein B modification: (SEP)467
Protein B sequence: RIGSGSFGTV
Protein B construct: RIGSG(pSer)FGTV (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.03
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): 2.74
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 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: BRAF
Protein B fragment: 334-343
Protein B site: 14-3-3_pSer339
Protein B modification: (SEP)339
Protein B sequence: PSPSKSIPIP
Protein B construct: PSPSK(pSer)IPIP (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.07
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): 2.32
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
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: BRAF
Protein B fragment: 609-618
Protein B site: 14-3-3_pSer614
Protein B modification: (SEP)614
Protein B sequence: QFEQLSGSIL
Protein B construct: QFEQL(pSer)GSIL (crude synthetic peptide)
Average holdup BI: -0.4
Holdup BI standard deviation: 0.32
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): 2.06
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: BRAF
Protein B fragment: 360-369
Protein B site: 14-3-3_pSer365
Protein B modification: (SEP)365
Protein B sequence: RDRSSSAPNV
Protein B construct: RDRSS(pSer)APNV (crude synthetic peptide)
Average holdup BI: 0.82
Holdup BI standard deviation: 0.18
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): 1.46
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: 4.5
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: BRAF
Protein B fragment: 462-471
Protein B site: 14-3-3_pSer467
Protein B modification: (SEP)467
Protein B sequence: RIGSGSFGTV
Protein B construct: RIGSG(pSer)FGTV (crude synthetic peptide)
Average holdup BI: 0.07
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): 1.33
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 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: BRAF
Protein B fragment: 724-733
Protein B site: 14-3-3_pSer729
Protein B modification: (SEP)729
Protein B sequence: IHRSASEPSL
Protein B construct: IHRSA(pSer)EPSL (crude synthetic peptide)
Average holdup BI: 0.69
Holdup BI standard deviation: 0.25
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): 1.33
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: 4.29
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: BRAF
Protein B fragment: 597-606
Protein B site: 14-3-3_pSer602
Protein B modification: (SEP)602
Protein B sequence: LATVKSRWSG
Protein B construct: LATVK(pSer)RWSG (crude synthetic peptide)
Average holdup BI: 0.05
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): 1.1
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.
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: BRAF
Protein B fragment: 441-450
Protein B site: 14-3-3_pSer446
Protein B modification: (SEP)446
Protein B sequence: LGRRDSSDDW
Protein B construct: LGRRD(pSer)SDDW (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.19
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): 0.83
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: BRAF
Protein B fragment: 594-603
Protein B site: 14-3-3_pThr599
Protein B modification: (TPO)599
Protein B sequence: DFGLATVKSR
Protein B construct: DFGLA(pThr)VKSR (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.07
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): 0.72
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 15
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: BRAF
Protein B fragment: 389-398
Protein B site: 14-3-3_pSer394
Protein B modification: (SEP)394
Protein B sequence: RGDGGSTTGL
Protein B construct: RGDGG(pSer)TTGL (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.08
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): 0.5
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 16
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: BRAF
Protein B fragment: 597-606
Protein B site: 14-3-3_pSer602
Protein B modification: (SEP)602
Protein B sequence: LATVKSRWSG
Protein B construct: LATVK(pSer)RWSG (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.07
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): 0.35
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 17
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: BRAF
Protein B fragment: 594-603
Protein B site: 14-3-3_pThr599
Protein B modification: (TPO)599
Protein B sequence: DFGLATVKSR
Protein B construct: DFGLA(pThr)VKSR (crude synthetic peptide)
Average holdup BI: 0.02
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): 0.26
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 18
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: BRAF
Protein B fragment: 609-618
Protein B site: 14-3-3_pSer614
Protein B modification: (SEP)614
Protein B sequence: QFEQLSGSIL
Protein B construct: QFEQL(pSer)GSIL (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.2
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): 0.21
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 19
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: BRAF
Protein B fragment: 389-398
Protein B site: 14-3-3_pSer394
Protein B modification: (SEP)394
Protein B sequence: RGDGGSTTGL
Protein B construct: RGDGG(pSer)TTGL (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.08
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): 0.2
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 20
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: BRAF
Protein B fragment: 574-583
Protein B site: 14-3-3_pSer579
Protein B modification: (SEP)579
Protein B sequence: HRDLKSNNIF
Protein B construct: HRDLK(pSer)NNIF (crude synthetic peptide)
Average holdup BI: -0.07
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): 0.12
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 21
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: BRAF
Protein B fragment: 574-583
Protein B site: 14-3-3_pSer579
Protein B modification: (SEP)579
Protein B sequence: HRDLKSNNIF
Protein B construct: HRDLK(pSer)NNIF (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.49
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): 0.07
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.
Experiment series 22
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: BRAF
Protein B fragment: 441-450
Protein B site: 14-3-3_pSer446
Protein B modification: (SEP)446
Protein B sequence: LGRRDSSDDW
Protein B construct: LGRRD(pSer)SDDW (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.1
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): 0.02
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.