Details of experiments between P31947 and Q86X27
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: RALGPS2
Protein B fragment: 356-365
Protein B site: 14-3-3_pThr361
Protein B modification: (TPO)361
Protein B sequence: EFKSATFPNA
Protein B construct: EFKSA(pThr)FPNA (crude synthetic peptide)
Average holdup BI: 0.87
Holdup BI standard deviation: 0.01
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): 10.11
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
Measured pKd: 4.76
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: RALGPS2
Protein B fragment: 356-365
Protein B site: 14-3-3_pThr361
Protein B modification: (TPO)361
Protein B sequence: EFKSATFPNA
Protein B construct: EFKSA(pThr)FPNA (crude synthetic peptide)
Average holdup BI: 0.84
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): 8.81
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: 4.68
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: RALGPS2
Protein B fragment: 356-365
Protein B site: 14-3-3_pThr361
Protein B modification: (TPO)361
Protein B sequence: EFKSATFPNA
Protein B construct: EFKSA(pThr)FPNA (crude synthetic peptide)
Average holdup BI: 0.9
Holdup BI standard deviation: 0.01
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): 8.78
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: 4.78
Experiment series 4
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: RALGPS2
Protein B fragment: 417-426
Protein B site: 14-3-3_pSer422
Protein B modification: (SEP)422
Protein B sequence: NRLYHSLGPV
Protein B construct: NRLYH(pSer)LGPV (crude synthetic peptide)
Average holdup BI: 0.65
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): 5.66
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: 4.22
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: RALGPS2
Protein B fragment: 417-426
Protein B site: 14-3-3_pSer422
Protein B modification: (SEP)422
Protein B sequence: NRLYHSLGPV
Protein B construct: NRLYH(pSer)LGPV (crude synthetic peptide)
Average holdup BI: 0.68
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): 5.58
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
Measured pKd: 4.16
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: RALGPS2
Protein B fragment: 417-426
Protein B site: 14-3-3_pSer422
Protein B modification: (SEP)422
Protein B sequence: NRLYHSLGPV
Protein B construct: NRLYH(pSer)LGPV (crude synthetic peptide)
Average holdup BI: 0.56
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): 4.69
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: 4.04
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: RALGPS2
Protein B fragment: 291-300
Protein B site: 14-3-3_pSer296
Protein B modification: (SEP)296
Protein B sequence: PRSAASREDL
Protein B construct: PRSAA(pSer)REDL (crude synthetic peptide)
Average holdup BI: -0.1
Holdup BI standard deviation: 0.15
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): 0.72
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
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: RALGPS2
Protein B fragment: 291-300
Protein B site: 14-3-3_pSer296
Protein B modification: (SEP)296
Protein B sequence: PRSAASREDL
Protein B construct: PRSAA(pSer)REDL (crude synthetic peptide)
Average holdup BI: -0.01
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): 0.11
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 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: RALGPS2
Protein B fragment: 291-300
Protein B site: 14-3-3_pSer296
Protein B modification: (SEP)296
Protein B sequence: PRSAASREDL
Protein B construct: PRSAA(pSer)REDL (crude synthetic peptide)
Average holdup BI: 0.01
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): 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.