Details of experiments between P31946 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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.38
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.41
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: 3.78
Experiment series 2
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.53
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.13
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
Measured pKd: 3.93
Experiment series 3
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.45
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.26
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: 3.91
Experiment series 4
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.38
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.99
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: -0.06
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.9
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 6
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.06
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.01
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.11
Holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.92
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.04
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.82
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: -0.08
Holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.71
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.11
Holdup BI standard deviation: 0.23
Immobilized partner concentration in holdup experiment (10-6M): 102
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: 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 11
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: -0.48
Holdup BI standard deviation: 1.75
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.37
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.23
Holdup BI standard deviation: 0.52
Immobilized partner concentration in holdup experiment (10-6M): 134
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: 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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.21
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.09
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.03
Holdup BI standard deviation: 0.27
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.07
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 102
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: 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.