Details of experiments between P31947 and O94915
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: FRYL
Protein B fragment: 1954-1963
Protein B site: 14-3-3_pThr1959
Protein B modification: (TPO)1959
Protein B sequence: RRRSNTLDIM
Protein B construct: RRRSN(pThr)LDIM (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 111
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: 1
Measured pKd: 4
Experiment series 2
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: FRYL
Protein B fragment: 1909-1918
Protein B site: 14-3-3_pSer1914
Protein B modification: (SEP)1914
Protein B sequence: AANRKSTGQL
Protein B construct: AANRK(pSer)TGQL (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 142
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: 1
Measured pKd: 4
Experiment series 3
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: FRYL
Protein B fragment: 1973-1982
Protein B site: 14-3-3_pSer1978
Protein B modification: (SEP)1978
Protein B sequence: LARTRSLSSL
Protein B construct: LARTR(pSer)LSSL (crude synthetic peptide)
Average holdup BI: 0.75
Holdup BI standard deviation: 0.03
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): 8.1
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: 4.32
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: FRYL
Protein B fragment: 1936-1945
Protein B site: 14-3-3_pSer1941
Protein B modification: (SEP)1941
Protein B sequence: NARSNSLRLS
Protein B construct: NARSN(pSer)LRLS (crude synthetic peptide)
Average holdup BI: 0.35
Holdup BI standard deviation: 0.02
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): 6.66
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: 3.68
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: FRYL
Protein B fragment: 1936-1945
Protein B site: 14-3-3_pSer1941
Protein B modification: (SEP)1941
Protein B sequence: NARSNSLRLS
Protein B construct: NARSN(pSer)LRLS (crude synthetic peptide)
Average holdup BI: 0.39
Holdup BI standard deviation: 0.03
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): 6.62
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: 3.65
Experiment series 6
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: FRYL
Protein B fragment: 1973-1982
Protein B site: 14-3-3_pSer1978
Protein B modification: (SEP)1978
Protein B sequence: LARTRSLSSL
Protein B construct: LARTR(pSer)LSSL (crude synthetic peptide)
Average holdup BI: 0.7
Holdup BI standard deviation: 0.03
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): 6.16
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
Measured pKd: 4.32
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: FRYL
Protein B fragment: 1973-1982
Protein B site: 14-3-3_pSer1978
Protein B modification: (SEP)1978
Protein B sequence: LARTRSLSSL
Protein B construct: LARTR(pSer)LSSL (crude synthetic peptide)
Average holdup BI: 0.69
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): 5.13
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
Measured pKd: 4.3
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: FRYL
Protein B fragment: 1936-1945
Protein B site: 14-3-3_pSer1941
Protein B modification: (SEP)1941
Protein B sequence: NARSNSLRLS
Protein B construct: NARSN(pSer)LRLS (crude synthetic peptide)
Average holdup BI: 0.31
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): 4.64
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: 3.6
Experiment series 9
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: FRYL
Protein B fragment: 1954-1963
Protein B site: 14-3-3_pThr1959
Protein B modification: (TPO)1959
Protein B sequence: RRRSNTLDIM
Protein B construct: RRRSN(pThr)LDIM (crude synthetic peptide)
Average holdup BI: 0.57
Holdup BI standard deviation: 0.13
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): 3.34
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
Measured pKd: 3.97
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: FRYL
Protein B fragment: 1954-1963
Protein B site: 14-3-3_pThr1959
Protein B modification: (TPO)1959
Protein B sequence: RRRSNTLDIM
Protein B construct: RRRSN(pThr)LDIM (crude synthetic peptide)
Average holdup BI: 0.6
Holdup BI standard deviation: 0.2
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): 1.63
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: 4.11
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: FRYL
Protein B fragment: 1930-1939
Protein B site: 14-3-3_pSer1935
Protein B modification: (SEP)1935
Protein B sequence: YLGYNSNARS
Protein B construct: YLGYN(pSer)NARS (crude synthetic peptide)
Average holdup BI: 0.06
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): 1.06
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 12
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: FRYL
Protein B fragment: 1967-1976
Protein B site: 14-3-3_pSer1972
Protein B modification: (SEP)1972
Protein B sequence: INHSSSLART
Protein B construct: INHSS(pSer)LART (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.1
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): 0.77
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: 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: FRYL
Protein B fragment: 477-486
Protein B site: 14-3-3_pThr482
Protein B modification: (TPO)482
Protein B sequence: LPSGNTLRVK
Protein B construct: LPSGN(pThr)LRVK (crude synthetic peptide)
Average holdup BI: 0.03
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): 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 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: FRYL
Protein B fragment: 98-107
Protein B site: 14-3-3_pSer103
Protein B modification: (SEP)103
Protein B sequence: RSSTKSKGDE
Protein B construct: RSSTK(pSer)KGDE (crude synthetic peptide)
Average holdup BI: -0.3
Holdup BI standard deviation: 0.56
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): 0.62
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: 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: FRYL
Protein B fragment: 477-486
Protein B site: 14-3-3_pThr482
Protein B modification: (TPO)482
Protein B sequence: LPSGNTLRVK
Protein B construct: LPSGN(pThr)LRVK (crude synthetic peptide)
Average holdup BI: 0.02
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): 0.61
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: FRYL
Protein B fragment: 1967-1976
Protein B site: 14-3-3_pSer1972
Protein B modification: (SEP)1972
Protein B sequence: INHSSSLART
Protein B construct: INHSS(pSer)LART (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.04
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): 0.55
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 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: FRYL
Protein B fragment: 98-107
Protein B site: 14-3-3_pSer103
Protein B modification: (SEP)103
Protein B sequence: RSSTKSKGDE
Protein B construct: RSSTK(pSer)KGDE (crude synthetic peptide)
Average holdup BI: -0.3
Holdup BI standard deviation: 0.57
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): 0.53
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: 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: FRYL
Protein B fragment: 1967-1976
Protein B site: 14-3-3_pSer1972
Protein B modification: (SEP)1972
Protein B sequence: INHSSSLART
Protein B construct: INHSS(pSer)LART (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.09
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): 0.51
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 19
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: FRYL
Protein B fragment: 1930-1939
Protein B site: 14-3-3_pSer1935
Protein B modification: (SEP)1935
Protein B sequence: YLGYNSNARS
Protein B construct: YLGYN(pSer)NARS (crude synthetic peptide)
Average holdup BI: 0.02
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): 0.38
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 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: FRYL
Protein B fragment: 1930-1939
Protein B site: 14-3-3_pSer1935
Protein B modification: (SEP)1935
Protein B sequence: YLGYNSNARS
Protein B construct: YLGYN(pSer)NARS (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.04
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): 0.23
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: FRYL
Protein B fragment: 1940-1949
Protein B site: 14-3-3_pSer1945
Protein B modification: (SEP)1945
Protein B sequence: NSLRLSLIGD
Protein B construct: NSLRL(pSer)LIGD (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.16
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): 0.23
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: 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: FRYL
Protein B fragment: 1909-1918
Protein B site: 14-3-3_pSer1914
Protein B modification: (SEP)1914
Protein B sequence: AANRKSTGQL
Protein B construct: AANRK(pSer)TGQL (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.16
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 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: FRYL
Protein B fragment: 477-486
Protein B site: 14-3-3_pThr482
Protein B modification: (TPO)482
Protein B sequence: LPSGNTLRVK
Protein B construct: LPSGN(pThr)LRVK (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.09
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): 0.09
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 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: FRYL
Protein B fragment: 1940-1949
Protein B site: 14-3-3_pSer1945
Protein B modification: (SEP)1945
Protein B sequence: NSLRLSLIGD
Protein B construct: NSLRL(pSer)LIGD (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.11
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): 0.01
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.