Details of experiments between P31947 and Q16875
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: PFKFB3
Protein B fragment: 473-482
Protein B site: 14-3-3_pSer478
Protein B modification: (SEP)478
Protein B sequence: KPRINSFEEH
Protein B construct: KPRIN(pSer)FEEH (crude synthetic peptide)
Average holdup BI: -0.05
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): 1.28
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 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: PFKFB3
Protein B fragment: 17-26
Protein B site: 14-3-3_pSer22
Protein B modification: (SEP)22
Protein B sequence: VDHRPSLPRS
Protein B construct: VDHRP(pSer)LPRS (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.06
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): 1.15
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 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: PFKFB3
Protein B fragment: 473-482
Protein B site: 14-3-3_pSer478
Protein B modification: (SEP)478
Protein B sequence: KPRINSFEEH
Protein B construct: KPRIN(pSer)FEEH (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.06
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): 1.13
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 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: PFKFB3
Protein B fragment: 17-26
Protein B site: 14-3-3_pSer22
Protein B modification: (SEP)22
Protein B sequence: VDHRPSLPRS
Protein B construct: VDHRP(pSer)LPRS (crude synthetic peptide)
Average holdup BI: -0.12
Holdup BI standard deviation: 0.13
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): 1.02
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 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: PFKFB3
Protein B fragment: 473-482
Protein B site: 14-3-3_pSer478
Protein B modification: (SEP)478
Protein B sequence: KPRINSFEEH
Protein B construct: KPRIN(pSer)FEEH (crude synthetic peptide)
Average holdup BI: -0.11
Holdup BI standard deviation: 0.12
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): 0.95
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 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: PFKFB3
Protein B fragment: 456-465
Protein B site: 14-3-3_pSer461
Protein B modification: (SEP)461
Protein B sequence: LMRRNSVTPL
Protein B construct: LMRRN(pSer)VTPL (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.18
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): 0.7
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
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: PFKFB3
Protein B fragment: 456-465
Protein B site: 14-3-3_pSer461
Protein B modification: (SEP)461
Protein B sequence: LMRRNSVTPL
Protein B construct: LMRRN(pSer)VTPL (crude synthetic peptide)
Average holdup BI: -0.29
Holdup BI standard deviation: 0.5
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): 0.62
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 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: PFKFB3
Protein B fragment: 456-465
Protein B site: 14-3-3_pSer461
Protein B modification: (SEP)461
Protein B sequence: LMRRNSVTPL
Protein B construct: LMRRN(pSer)VTPL (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.12
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: PFKFB3
Protein B fragment: 17-26
Protein B site: 14-3-3_pSer22
Protein B modification: (SEP)22
Protein B sequence: VDHRPSLPRS
Protein B construct: VDHRP(pSer)LPRS (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): 0.06
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.