Details of experiments between P27348 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: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.9
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.24
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: 4.86
Experiment series 2
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.35
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.67
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: 3.61
Experiment series 3
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.29
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.18
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: 3.5
Experiment series 4
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.66
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 5
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: -0.44
Holdup BI standard deviation: 0.33
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.26
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 6
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.92
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.69
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
Measured pKd: 4.96
Experiment series 7
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: -0.06
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.9
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: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: -0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.42
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: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: -0.03
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.34
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: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: BRAF
Protein B fragment: 600-609
Protein B site: 14-3-3_pSer605
Protein B modification: (SEP)605
Protein B sequence: VKSRWSGSHQ
Protein B construct: VKSRW(pSer)GSHQ (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.21
Immobilized partner concentration in holdup experiment (10-6M): 130
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: 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: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 130
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.
Experiment series 12
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: -0.01
Holdup BI standard deviation: 0.45
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.01
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.