Details of experiments between P31947 and Q96PY6
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: NEK1
Protein B fragment: 409-418
Protein B site: 14-3-3_pSer414
Protein B modification: (SEP)414
Protein B sequence: WRNVLSAGGS
Protein B construct: WRNVL(pSer)AGGS (crude synthetic peptide)
Average holdup BI: 0.29
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): 3.57
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: 3.56
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: NEK1
Protein B fragment: 409-418
Protein B site: 14-3-3_pSer414
Protein B modification: (SEP)414
Protein B sequence: WRNVLSAGGS
Protein B construct: WRNVL(pSer)AGGS (crude synthetic peptide)
Average holdup BI: 0.23
Holdup BI standard deviation: 0.08
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): 3.04
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: 3.33
Experiment series 3
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: NEK1
Protein B fragment: 409-418
Protein B site: 14-3-3_pSer414
Protein B modification: (SEP)414
Protein B sequence: WRNVLSAGGS
Protein B construct: WRNVL(pSer)AGGS (crude synthetic peptide)
Average holdup BI: 0.17
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): 2.88
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 construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: NEK1
Protein B fragment: 136-145
Protein B site: 14-3-3_pThr141
Protein B modification: (TPO)141
Protein B sequence: LTKDGTVQLG
Protein B construct: LTKDG(pThr)VQLG (crude synthetic peptide)
Average holdup BI: 0.03
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): 0.91
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: NEK1
Protein B fragment: 9-18
Protein B site: 14-3-3_pSer14
Protein B modification: (SEP)14
Protein B sequence: KIGEGSFGKA
Protein B construct: KIGEG(pSer)FGKA (crude synthetic peptide)
Average holdup BI: -0.04
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): 0.8
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 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: NEK1
Protein B fragment: 280-289
Protein B site: 14-3-3_pSer285
Protein B modification: (SEP)285
Protein B sequence: FSKFGSQPIP
Protein B construct: FSKFG(pSer)QPIP (crude synthetic peptide)
Average holdup BI: 0.2
Holdup BI standard deviation: 0.27
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): 0.74
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 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: NEK1
Protein B fragment: 294-303
Protein B site: 14-3-3_pSer299
Protein B modification: (SEP)299
Protein B sequence: ASGQNSISVM
Protein B construct: ASGQN(pSer)ISVM (crude synthetic peptide)
Average holdup BI: 0.12
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): 0.66
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 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: NEK1
Protein B fragment: 151-160
Protein B site: 14-3-3_pThr156
Protein B modification: (TPO)156
Protein B sequence: RVLNSTVELA
Protein B construct: RVLNS(pThr)VELA (crude synthetic peptide)
Average holdup BI: -0.04
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): 0.65
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 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: NEK1
Protein B fragment: 136-145
Protein B site: 14-3-3_pThr141
Protein B modification: (TPO)141
Protein B sequence: LTKDGTVQLG
Protein B construct: LTKDG(pThr)VQLG (crude synthetic peptide)
Average holdup BI: 0.03
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): 0.56
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
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: NEK1
Protein B fragment: 280-289
Protein B site: 14-3-3_pSer285
Protein B modification: (SEP)285
Protein B sequence: FSKFGSQPIP
Protein B construct: FSKFG(pSer)QPIP (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.2
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): 0.55
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: NEK1
Protein B fragment: 294-303
Protein B site: 14-3-3_pSer299
Protein B modification: (SEP)299
Protein B sequence: ASGQNSISVM
Protein B construct: ASGQN(pSer)ISVM (crude synthetic peptide)
Average holdup BI: -0.28
Holdup BI standard deviation: 0.51
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 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: NEK1
Protein B fragment: 413-422
Protein B site: 14-3-3_pSer418
Protein B modification: (SEP)418
Protein B sequence: LSAGGSGEVK
Protein B construct: LSAGG(pSer)GEVK (crude synthetic peptide)
Average holdup BI: -0.03
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): 0.45
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: NEK1
Protein B fragment: 151-160
Protein B site: 14-3-3_pThr156
Protein B modification: (TPO)156
Protein B sequence: RVLNSTVELA
Protein B construct: RVLNS(pThr)VELA (crude synthetic peptide)
Average holdup BI: -0.03
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): 0.42
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 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: NEK1
Protein B fragment: 151-160
Protein B site: 14-3-3_pThr156
Protein B modification: (TPO)156
Protein B sequence: RVLNSTVELA
Protein B construct: RVLNS(pThr)VELA (crude synthetic peptide)
Average holdup BI: 0.02
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): 0.33
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
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: NEK1
Protein B fragment: 294-303
Protein B site: 14-3-3_pSer299
Protein B modification: (SEP)299
Protein B sequence: ASGQNSISVM
Protein B construct: ASGQN(pSer)ISVM (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 0.51
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): 0.24
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
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: NEK1
Protein B fragment: 136-145
Protein B site: 14-3-3_pThr141
Protein B modification: (TPO)141
Protein B sequence: LTKDGTVQLG
Protein B construct: LTKDG(pThr)VQLG (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.04
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): 0.22
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 17
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: NEK1
Protein B fragment: 1047-1056
Protein B site: 14-3-3_pSer1052
Protein B modification: (SEP)1052
Protein B sequence: MLRTCSLPDL
Protein B construct: MLRTC(pSer)LPDL (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.31
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): 0.18
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 18
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: NEK1
Protein B fragment: 9-18
Protein B site: 14-3-3_pSer14
Protein B modification: (SEP)14
Protein B sequence: KIGEGSFGKA
Protein B construct: KIGEG(pSer)FGKA (crude synthetic peptide)
Average holdup BI: -0.01
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): 0.17
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 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: NEK1
Protein B fragment: 1047-1056
Protein B site: 14-3-3_pSer1052
Protein B modification: (SEP)1052
Protein B sequence: MLRTCSLPDL
Protein B construct: MLRTC(pSer)LPDL (crude synthetic peptide)
Average holdup BI: 0.21
Holdup BI standard deviation: 0.69
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): 0.17
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 construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: NEK1
Protein B fragment: 413-422
Protein B site: 14-3-3_pSer418
Protein B modification: (SEP)418
Protein B sequence: LSAGGSGEVK
Protein B construct: LSAGG(pSer)GEVK (crude synthetic peptide)
Average holdup BI: -0.01
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): 0.16
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 21
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: NEK1
Protein B fragment: 1121-1130
Protein B site: 14-3-3_pSer1126
Protein B modification: (SEP)1126
Protein B sequence: PGEEYSEEEE
Protein B construct: PGEEY(pSer)EEEE (crude synthetic peptide)
Average holdup BI: -0.47
Holdup BI standard deviation: 2.62
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): 0.15
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 22
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: NEK1
Protein B fragment: 1121-1130
Protein B site: 14-3-3_pSer1126
Protein B modification: (SEP)1126
Protein B sequence: PGEEYSEEEE
Protein B construct: PGEEY(pSer)EEEE (crude synthetic peptide)
Average holdup BI: -0.26
Holdup BI standard deviation: 1.67
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): 0.15
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 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: NEK1
Protein B fragment: 9-18
Protein B site: 14-3-3_pSer14
Protein B modification: (SEP)14
Protein B sequence: KIGEGSFGKA
Protein B construct: KIGEG(pSer)FGKA (crude synthetic peptide)
Average holdup BI: -0.01
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): 0.14
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: NEK1
Protein B fragment: 1047-1056
Protein B site: 14-3-3_pSer1052
Protein B modification: (SEP)1052
Protein B sequence: MLRTCSLPDL
Protein B construct: MLRTC(pSer)LPDL (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.65
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): 0.06
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 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: NEK1
Protein B fragment: 413-422
Protein B site: 14-3-3_pSer418
Protein B modification: (SEP)418
Protein B sequence: LSAGGSGEVK
Protein B construct: LSAGG(pSer)GEVK (crude synthetic peptide)
Average holdup BI: 0
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): 0
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.