Details of experiments between P31947 and Q92934
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: BAD
Protein B fragment: 94-103
Protein B site: 14-3-3_pSer99
Protein B modification: (SEP)99
Protein B sequence: RGRSRSAPPN
Protein B construct: RGRSR(pSer)APPN (crude synthetic peptide)
Average holdup BI: 0.51
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): 5.7
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: 3.96
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: BAD
Protein B fragment: 129-138
Protein B site: 14-3-3_pSer134
Protein B modification: (SEP)134
Protein B sequence: LPRPKSAGTA
Protein B construct: LPRPK(pSer)AGTA (crude synthetic peptide)
Average holdup BI: 0.51
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): 4.95
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: BAD
Protein B fragment: 129-138
Protein B site: 14-3-3_pSer134
Protein B modification: (SEP)134
Protein B sequence: LPRPKSAGTA
Protein B construct: LPRPK(pSer)AGTA (crude synthetic peptide)
Average holdup BI: 0.49
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): 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: 3.94
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: BAD
Protein B fragment: 94-103
Protein B site: 14-3-3_pSer99
Protein B modification: (SEP)99
Protein B sequence: RGRSRSAPPN
Protein B construct: RGRSR(pSer)APPN (crude synthetic peptide)
Average holdup BI: 0.34
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): 3.33
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
Measured pKd: 3.66
Experiment series 5
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: BAD
Protein B fragment: 129-138
Protein B site: 14-3-3_pSer134
Protein B modification: (SEP)134
Protein B sequence: LPRPKSAGTA
Protein B construct: LPRPK(pSer)AGTA (crude synthetic peptide)
Average holdup BI: 0.44
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): 3.33
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
Measured pKd: 3.85
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: BAD
Protein B fragment: 66-75
Protein B site: 14-3-3_pSer71
Protein B modification: (SEP)71
Protein B sequence: AVEIRSRHSS
Protein B construct: AVEIR(pSer)RHSS (crude synthetic peptide)
Average holdup BI: 0.11
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): 0.46
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 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: BAD
Protein B fragment: 113-122
Protein B site: 14-3-3_pSer118
Protein B modification: (SEP)118
Protein B sequence: ELRRMSDEFV
Protein B construct: ELRRM(pSer)DEFV (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.22
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): 0.43
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 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: BAD
Protein B fragment: 66-75
Protein B site: 14-3-3_pSer71
Protein B modification: (SEP)71
Protein B sequence: AVEIRSRHSS
Protein B construct: AVEIR(pSer)RHSS (crude synthetic peptide)
Average holdup BI: -0.22
Holdup BI standard deviation: 0.36
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): 0.34
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
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: BAD
Protein B fragment: 119-128
Protein B site: 14-3-3_pSer124
Protein B modification: (SEP)124
Protein B sequence: DEFVDSFKKG
Protein B construct: DEFVD(pSer)FKKG (crude synthetic peptide)
Average holdup BI: -0.02
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): 0.3
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: BAD
Protein B fragment: 66-75
Protein B site: 14-3-3_pSer71
Protein B modification: (SEP)71
Protein B sequence: AVEIRSRHSS
Protein B construct: AVEIR(pSer)RHSS (crude synthetic peptide)
Average holdup BI: 0.21
Holdup BI standard deviation: 0.55
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 11
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: BAD
Protein B fragment: 113-122
Protein B site: 14-3-3_pSer118
Protein B modification: (SEP)118
Protein B sequence: ELRRMSDEFV
Protein B construct: ELRRM(pSer)DEFV (crude synthetic peptide)
Average holdup BI: -0.1
Holdup BI standard deviation: 0.32
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): 0.25
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: BAD
Protein B fragment: 70-79
Protein B site: 14-3-3_pSer75
Protein B modification: (SEP)75
Protein B sequence: RSRHSSYPAG
Protein B construct: RSRHS(pSer)YPAG (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.68
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): 0.18
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 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: BAD
Protein B fragment: 119-128
Protein B site: 14-3-3_pSer124
Protein B modification: (SEP)124
Protein B sequence: DEFVDSFKKG
Protein B construct: DEFVD(pSer)FKKG (crude synthetic peptide)
Average holdup BI: 0
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): 0.1
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
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: BAD
Protein B fragment: 113-122
Protein B site: 14-3-3_pSer118
Protein B modification: (SEP)118
Protein B sequence: ELRRMSDEFV
Protein B construct: ELRRM(pSer)DEFV (crude synthetic peptide)
Average holdup BI: 0
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): 0.02
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: BAD
Protein B fragment: 119-128
Protein B site: 14-3-3_pSer124
Protein B modification: (SEP)124
Protein B sequence: DEFVDSFKKG
Protein B construct: DEFVD(pSer)FKKG (crude synthetic peptide)
Average holdup BI: 0
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): 0.02
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.