Details of experiments between P31947 and Q5VZ89
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: DENND4C
Protein B fragment: 1320-1329
Protein B site: 14-3-3_pSer1325
Protein B modification: (SEP)1325
Protein B sequence: KERSTSLSAL
Protein B construct: KERST(pSer)LSAL (crude synthetic peptide)
Average holdup BI: 0.5
Holdup BI standard deviation: 0.01
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): 7.92
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: 3.96
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: DENND4C
Protein B fragment: 1657-1666
Protein B site: 14-3-3_pSer1662
Protein B modification: (SEP)1662
Protein B sequence: GISTVSLPNS
Protein B construct: GISTV(pSer)LPNS (crude synthetic peptide)
Average holdup BI: 0.51
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): 6.82
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: 3.86
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: DENND4C
Protein B fragment: 1657-1666
Protein B site: 14-3-3_pSer1662
Protein B modification: (SEP)1662
Protein B sequence: GISTVSLPNS
Protein B construct: GISTV(pSer)LPNS (crude synthetic peptide)
Average holdup BI: 0.47
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): 6.8
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: 3.89
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: DENND4C
Protein B fragment: 1320-1329
Protein B site: 14-3-3_pSer1325
Protein B modification: (SEP)1325
Protein B sequence: KERSTSLSAL
Protein B construct: KERST(pSer)LSAL (crude synthetic peptide)
Average holdup BI: 0.59
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): 6.51
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
Measured pKd: 4
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: DENND4C
Protein B fragment: 1320-1329
Protein B site: 14-3-3_pSer1325
Protein B modification: (SEP)1325
Protein B sequence: KERSTSLSAL
Protein B construct: KERST(pSer)LSAL (crude synthetic peptide)
Average holdup BI: 0.51
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): 6.1
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
Measured pKd: 3.97
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: DENND4C
Protein B fragment: 1657-1666
Protein B site: 14-3-3_pSer1662
Protein B modification: (SEP)1662
Protein B sequence: GISTVSLPNS
Protein B construct: GISTV(pSer)LPNS (crude synthetic peptide)
Average holdup BI: 0.42
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): 5.69
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: 3.81
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: DENND4C
Protein B fragment: 1635-1644
Protein B site: 14-3-3_pSer1640
Protein B modification: (SEP)1640
Protein B sequence: LTRSHSVGGP
Protein B construct: LTRSH(pSer)VGGP (crude synthetic peptide)
Average holdup BI: 0.51
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): 5.56
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: 3.87
Experiment series 8
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: DENND4C
Protein B fragment: 1618-1627
Protein B site: 14-3-3_pSer1623
Protein B modification: (SEP)1623
Protein B sequence: TMKISSVPNS
Protein B construct: TMKIS(pSer)VPNS (crude synthetic peptide)
Average holdup BI: 0.26
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): 5.25
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: 3.4
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: DENND4C
Protein B fragment: 1273-1282
Protein B site: 14-3-3_pSer1278
Protein B modification: (SEP)1278
Protein B sequence: LERRSSLPLD
Protein B construct: LERRS(pSer)LPLD (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): 4.25
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: 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: DENND4C
Protein B fragment: 1635-1644
Protein B site: 14-3-3_pSer1640
Protein B modification: (SEP)1640
Protein B sequence: LTRSHSVGGP
Protein B construct: LTRSH(pSer)VGGP (crude synthetic peptide)
Average holdup BI: 0.37
Holdup BI standard deviation: 0.09
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): 3.4
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: 3.71
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: DENND4C
Protein B fragment: 1273-1282
Protein B site: 14-3-3_pSer1278
Protein B modification: (SEP)1278
Protein B sequence: LERRSSLPLD
Protein B construct: LERRS(pSer)LPLD (crude synthetic peptide)
Average holdup BI: 0.14
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): 3.13
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 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: DENND4C
Protein B fragment: 948-957
Protein B site: 14-3-3_pSer953
Protein B modification: (SEP)953
Protein B sequence: LIRLESIDNH
Protein B construct: LIRLE(pSer)IDNH (crude synthetic peptide)
Average holdup BI: -0.21
Holdup BI standard deviation: 0.08
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): 2.99
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 construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: DENND4C
Protein B fragment: 1618-1627
Protein B site: 14-3-3_pSer1623
Protein B modification: (SEP)1623
Protein B sequence: TMKISSVPNS
Protein B construct: TMKIS(pSer)VPNS (crude synthetic peptide)
Average holdup BI: 0.31
Holdup BI standard deviation: 0.11
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): 2.62
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
Measured pKd: 3.61
Experiment series 14
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: DENND4C
Protein B fragment: 1618-1627
Protein B site: 14-3-3_pSer1623
Protein B modification: (SEP)1623
Protein B sequence: TMKISSVPNS
Protein B construct: TMKIS(pSer)VPNS (crude synthetic peptide)
Average holdup BI: 0.34
Holdup BI standard deviation: 0.16
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): 2.24
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: 3.66
Experiment series 15
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: DENND4C
Protein B fragment: 1273-1282
Protein B site: 14-3-3_pSer1278
Protein B modification: (SEP)1278
Protein B sequence: LERRSSLPLD
Protein B construct: LERRS(pSer)LPLD (crude synthetic peptide)
Average holdup BI: 0.13
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): 1.71
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: DENND4C
Protein B fragment: 948-957
Protein B site: 14-3-3_pSer953
Protein B modification: (SEP)953
Protein B sequence: LIRLESIDNH
Protein B construct: LIRLE(pSer)IDNH (crude synthetic peptide)
Average holdup BI: -0.12
Holdup BI standard deviation: 0.1
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): 1.67
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 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: DENND4C
Protein B fragment: 1341-1350
Protein B site: 14-3-3_pSer1346
Protein B modification: (SEP)1346
Protein B sequence: VVNSLSGLKL
Protein B construct: VVNSL(pSer)GLKL (crude synthetic peptide)
Average holdup BI: -0.27
Holdup BI standard deviation: 0.26
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): 1.61
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: DENND4C
Protein B fragment: 1341-1350
Protein B site: 14-3-3_pSer1346
Protein B modification: (SEP)1346
Protein B sequence: VVNSLSGLKL
Protein B construct: VVNSL(pSer)GLKL (crude synthetic peptide)
Average holdup BI: 0.1
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): 1.15
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: DENND4C
Protein B fragment: 960-969
Protein B site: 14-3-3_pSer965
Protein B modification: (SEP)965
Protein B sequence: IVAKHSQPSP
Protein B construct: IVAKH(pSer)QPSP (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.11
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): 0.84
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: DENND4C
Protein B fragment: 948-957
Protein B site: 14-3-3_pSer953
Protein B modification: (SEP)953
Protein B sequence: LIRLESIDNH
Protein B construct: LIRLE(pSer)IDNH (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.06
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): 0.8
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: DENND4C
Protein B fragment: 960-969
Protein B site: 14-3-3_pSer965
Protein B modification: (SEP)965
Protein B sequence: IVAKHSQPSP
Protein B construct: IVAKH(pSer)QPSP (crude synthetic peptide)
Average holdup BI: -0.1
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): 0.75
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: DENND4C
Protein B fragment: 1179-1188
Protein B site: 14-3-3_pSer1184
Protein B modification: (SEP)1184
Protein B sequence: RNKRSSLYGI
Protein B construct: RNKRS(pSer)LYGI (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.19
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): 0.69
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: DENND4C
Protein B fragment: 1094-1103
Protein B site: 14-3-3_pSer1099
Protein B modification: (SEP)1099
Protein B sequence: IARTHSFENV
Protein B construct: IARTH(pSer)FENV (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.08
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 24
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: DENND4C
Protein B fragment: 1094-1103
Protein B site: 14-3-3_pSer1099
Protein B modification: (SEP)1099
Protein B sequence: IARTHSFENV
Protein B construct: IARTH(pSer)FENV (crude synthetic peptide)
Average holdup BI: 0.03
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): 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 25
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: DENND4C
Protein B fragment: 960-969
Protein B site: 14-3-3_pSer965
Protein B modification: (SEP)965
Protein B sequence: IVAKHSQPSP
Protein B construct: IVAKH(pSer)QPSP (crude synthetic peptide)
Average holdup BI: -0.04
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): 0.46
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 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: DENND4C
Protein B fragment: 1179-1188
Protein B site: 14-3-3_pSer1184
Protein B modification: (SEP)1184
Protein B sequence: RNKRSSLYGI
Protein B construct: RNKRS(pSer)LYGI (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.14
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): 0.46
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
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: DENND4C
Protein B fragment: 984-993
Protein B site: 14-3-3_pSer989
Protein B modification: (SEP)989
Protein B sequence: IVKVPSGIFD
Protein B construct: IVKVP(pSer)GIFD (crude synthetic peptide)
Average holdup BI: -0.19
Holdup BI standard deviation: 0.37
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): 0.46
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: DENND4C
Protein B fragment: 984-993
Protein B site: 14-3-3_pSer989
Protein B modification: (SEP)989
Protein B sequence: IVKVPSGIFD
Protein B construct: IVKVP(pSer)GIFD (crude synthetic peptide)
Average holdup BI: -0.05
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): 0.43
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 construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: DENND4C
Protein B fragment: 1041-1050
Protein B site: 14-3-3_pSer1046
Protein B modification: (SEP)1046
Protein B sequence: PERSCSFSSE
Protein B construct: PERSC(pSer)FSSE (crude synthetic peptide)
Average holdup BI: -0.47
Holdup BI standard deviation: 1.27
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): 0.39
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: DENND4C
Protein B fragment: 1661-1670
Protein B site: 14-3-3_pSer1666
Protein B modification: (SEP)1666
Protein B sequence: VSLPNSLQEV
Protein B construct: VSLPN(pSer)LQEV (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.37
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): 0.34
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: DENND4C
Protein B fragment: 1041-1050
Protein B site: 14-3-3_pSer1046
Protein B modification: (SEP)1046
Protein B sequence: PERSCSFSSE
Protein B construct: PERSC(pSer)FSSE (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.22
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): 0.29
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 32
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: DENND4C
Protein B fragment: 1179-1188
Protein B site: 14-3-3_pSer1184
Protein B modification: (SEP)1184
Protein B sequence: RNKRSSLYGI
Protein B construct: RNKRS(pSer)LYGI (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.08
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): 0.17
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 33
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: DENND4C
Protein B fragment: 1661-1670
Protein B site: 14-3-3_pSer1666
Protein B modification: (SEP)1666
Protein B sequence: VSLPNSLQEV
Protein B construct: VSLPN(pSer)LQEV (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.18
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): 0.16
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
The affinity was below the detection threshold of the assay.
Experiment series 34
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: DENND4C
Protein B fragment: 1661-1670
Protein B site: 14-3-3_pSer1666
Protein B modification: (SEP)1666
Protein B sequence: VSLPNSLQEV
Protein B construct: VSLPN(pSer)LQEV (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.46
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): 0.09
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
The affinity was below the detection threshold of the assay.
Experiment series 35
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: DENND4C
Protein B fragment: 1341-1350
Protein B site: 14-3-3_pSer1346
Protein B modification: (SEP)1346
Protein B sequence: VVNSLSGLKL
Protein B construct: VVNSL(pSer)GLKL (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.21
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): 0.08
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 36
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: DENND4C
Protein B fragment: 1094-1103
Protein B site: 14-3-3_pSer1099
Protein B modification: (SEP)1099
Protein B sequence: IARTHSFENV
Protein B construct: IARTH(pSer)FENV (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.14
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): 0.06
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 37
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: DENND4C
Protein B fragment: 1320-1329
Protein B site: 14-3-3_pSer1325
Protein B modification: (SEP)1325
Protein B sequence: KERSTSLSAL
Protein B construct: biotin-ttds-KERST(pSer)LSAL (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 132
Standard deviation of dissociation constant (10-6M): 14
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022.02.09
Number of measurements: 3
Measured pKd: 3.88