Details of experiments between P31947 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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.73
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.39
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: 4.28


Experiment series 2

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.37
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.7
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: 3.73


Experiment series 3

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.61
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.83
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
Measured pKd: 4.15


Experiment series 4

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.27
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.72
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: 3.42


Experiment series 5

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.19
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.45
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.14
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.74
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.78
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.6
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: 4.48


Experiment series 8

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.22
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.57
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: 3.41


Experiment series 9

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.13
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.04
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 10

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.22
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.72
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
Measured pKd: 3.38


Experiment series 11

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 113
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: 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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.88
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 142
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: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.7


Experiment series 13

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.19
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.08
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.73
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
The affinity was below the detection threshold of the assay.


Experiment series 15

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.26
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 16

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.42
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.2
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 17

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.06
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.03
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 18

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.07
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.86
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.04
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.67
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
The affinity was below the detection threshold of the assay.


Experiment series 20

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 113
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: 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 21

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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): 142
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: 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 22

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.33
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 23

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.06
Holdup BI standard deviation: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.33
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
The affinity was below the detection threshold of the assay.


Experiment series 24

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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): 111
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 25

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.24
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
The affinity was below the detection threshold of the assay.


Experiment series 26

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.12
Holdup BI standard deviation: 0.54
Immobilized partner concentration in holdup experiment (10-6M): 113
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: 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 27

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.18
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 28

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.13
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 29

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.1
Holdup BI standard deviation: 0.74
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.11
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 30

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.08
Holdup BI standard deviation: 0.51
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.11
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 31

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.06
Holdup BI standard deviation: 0.48
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.11
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 32

Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.01
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 111
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.