Details of experiments between P31947 and Q9H4L5
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: OSBPL3
Protein B fragment: 246-255
Protein B site: 14-3-3_pSer251
Protein B modification: (SEP)251
Protein B sequence: LHRTYSAPAI
Protein B construct: LHRTY(pSer)APAI (crude synthetic peptide)
Average holdup BI: 0.67
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): 6.26
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
Measured pKd: 4.15
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: OSBPL3
Protein B fragment: 299-308
Protein B site: 14-3-3_pSer304
Protein B modification: (SEP)304
Protein B sequence: VRLHSSNPNL
Protein B construct: VRLHS(pSer)NPNL (crude synthetic peptide)
Average holdup BI: 0.16
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): 5.08
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 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: OSBPL3
Protein B fragment: 299-308
Protein B site: 14-3-3_pSer304
Protein B modification: (SEP)304
Protein B sequence: VRLHSSNPNL
Protein B construct: VRLHS(pSer)NPNL (crude synthetic peptide)
Average holdup BI: 0.21
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): 4.16
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: 3.37
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: OSBPL3
Protein B fragment: 299-308
Protein B site: 14-3-3_pSer304
Protein B modification: (SEP)304
Protein B sequence: VRLHSSNPNL
Protein B construct: VRLHS(pSer)NPNL (crude synthetic peptide)
Average holdup BI: 0.27
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): 3.69
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: 3.41
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: OSBPL3
Protein B fragment: 246-255
Protein B site: 14-3-3_pSer251
Protein B modification: (SEP)251
Protein B sequence: LHRTYSAPAI
Protein B construct: LHRTY(pSer)APAI (crude synthetic peptide)
Average holdup BI: 0.81
Holdup BI standard deviation: 0.1
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): 2.6
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: 4.58
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: OSBPL3
Protein B fragment: 315-324
Protein B site: 14-3-3_pSer320
Protein B modification: (SEP)320
Protein B sequence: EEKNYSDGSE
Protein B construct: EEKNY(pSer)DGSE (crude synthetic peptide)
Average holdup BI: 0.93
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): 1.41
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: 5.06
Experiment series 7
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: OSBPL3
Protein B fragment: 432-441
Protein B site: 14-3-3_pSer437
Protein B modification: (SEP)437
Protein B sequence: LVHQLSNESR
Protein B construct: LVHQL(pSer)NESR (crude synthetic peptide)
Average holdup BI: -0.06
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): 0.54
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: 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: OSBPL3
Protein B fragment: 29-38
Protein B site: 14-3-3_pSer34
Protein B modification: (SEP)34
Protein B sequence: GSRQDSWEVV
Protein B construct: GSRQD(pSer)WEVV (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): 0.5
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: 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: OSBPL3
Protein B fragment: 29-38
Protein B site: 14-3-3_pSer34
Protein B modification: (SEP)34
Protein B sequence: GSRQDSWEVV
Protein B construct: GSRQD(pSer)WEVV (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.13
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 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: OSBPL3
Protein B fragment: 432-441
Protein B site: 14-3-3_pSer437
Protein B modification: (SEP)437
Protein B sequence: LVHQLSNESR
Protein B construct: LVHQL(pSer)NESR (crude synthetic peptide)
Average holdup BI: -0.01
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): 0.11
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 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: OSBPL3
Protein B fragment: 432-441
Protein B site: 14-3-3_pSer437
Protein B modification: (SEP)437
Protein B sequence: LVHQLSNESR
Protein B construct: LVHQL(pSer)NESR (crude synthetic peptide)
Average holdup BI: 0
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): 0.1
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
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: OSBPL3
Protein B fragment: 29-38
Protein B site: 14-3-3_pSer34
Protein B modification: (SEP)34
Protein B sequence: GSRQDSWEVV
Protein B construct: GSRQD(pSer)WEVV (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.26
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): 0.03
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.