Details of experiments between P62258 and Q8TF72
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 414-423
Protein B site: 14-3-3_pSer419
Protein B modification: (SEP)419
Protein B sequence: RPQANSLGSL
Protein B construct: RPQAN(pSer)LGSL (crude synthetic peptide)
Average holdup BI: 0.49
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.36
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: 3.92
Experiment series 2
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 414-423
Protein B site: 14-3-3_pSer419
Protein B modification: (SEP)419
Protein B sequence: RPQANSLGSL
Protein B construct: RPQAN(pSer)LGSL (crude synthetic peptide)
Average holdup BI: 0.55
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.84
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: 3.94
Experiment series 3
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 1457-1466
Protein B site: 14-3-3_pSer1462
Protein B modification: (SEP)1462
Protein B sequence: YQKQQSLPSL
Protein B construct: YQKQQ(pSer)LPSL (crude synthetic peptide)
Average holdup BI: 0.66
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.74
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
Measured pKd: 4.13
Experiment series 4
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 618-627
Protein B site: 14-3-3_pSer623
Protein B modification: (SEP)623
Protein B sequence: RSSRLSEPWE
Protein B construct: RSSRL(pSer)EPWE (crude synthetic peptide)
Average holdup BI: 0.28
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.21
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: 3.44
Experiment series 5
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 1416-1425
Protein B site: 14-3-3_pSer1421
Protein B modification: (SEP)1421
Protein B sequence: TRKRVSLPQW
Protein B construct: TRKRV(pSer)LPQW (crude synthetic peptide)
Average holdup BI: 0.76
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.66
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
Measured pKd: 4.35
Experiment series 6
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 618-627
Protein B site: 14-3-3_pSer623
Protein B modification: (SEP)623
Protein B sequence: RSSRLSEPWE
Protein B construct: RSSRL(pSer)EPWE (crude synthetic peptide)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.25
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 7
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 1216-1225
Protein B site: 14-3-3_pSer1221
Protein B modification: (SEP)1221
Protein B sequence: AGKSMSAEDL
Protein B construct: AGKSM(pSer)AEDL (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.7
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: 3.08
Experiment series 8
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 1216-1225
Protein B site: 14-3-3_pSer1221
Protein B modification: (SEP)1221
Protein B sequence: AGKSMSAEDL
Protein B construct: AGKSM(pSer)AEDL (crude synthetic peptide)
Average holdup BI: 0.26
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.76
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: 3.49
Experiment series 9
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 1016-1025
Protein B site: 14-3-3_pSer1021
Protein B modification: (SEP)1021
Protein B sequence: KKRSYSEPEK
Protein B construct: KKRSY(pSer)EPEK (crude synthetic peptide)
Average holdup BI: -0.1
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.3
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 107-116
Protein B site: 14-3-3_pThr112
Protein B modification: (TPO)112
Protein B sequence: RRDVCTDPGH
Protein B construct: RRDVC(pThr)DPGH (crude synthetic peptide)
Average holdup BI: -0.48
Holdup BI standard deviation: 0.65
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.08
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 1436-1445
Protein B site: 14-3-3_pSer1441
Protein B modification: (SEP)1441
Protein B sequence: AAREDSLPEE
Protein B construct: AARED(pSer)LPEE (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.83
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 716-725
Protein B site: 14-3-3_pSer721
Protein B modification: (SEP)721
Protein B sequence: LDRQVSYPRP
Protein B construct: LDRQV(pSer)YPRP (crude synthetic peptide)
Average holdup BI: -0.21
Holdup BI standard deviation: 0.39
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.57
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
The affinity was below the detection threshold of the assay.
Experiment series 13
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 949-958
Protein B site: 14-3-3_pSer954
Protein B modification: (SEP)954
Protein B sequence: GAPVASRSWR
Protein B construct: GAPVA(pSer)RSWR (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.38
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 716-725
Protein B site: 14-3-3_pSer721
Protein B modification: (SEP)721
Protein B sequence: LDRQVSYPRP
Protein B construct: LDRQV(pSer)YPRP (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.35
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 1436-1445
Protein B site: 14-3-3_pSer1441
Protein B modification: (SEP)1441
Protein B sequence: AAREDSLPEE
Protein B construct: AARED(pSer)LPEE (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 116
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 16
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 107-116
Protein B site: 14-3-3_pThr112
Protein B modification: (TPO)112
Protein B sequence: RRDVCTDPGH
Protein B construct: RRDVC(pThr)DPGH (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 143
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 17
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 1416-1425
Protein B site: 14-3-3_pSer1421
Protein B modification: (SEP)1421
Protein B sequence: TRKRVSLPQW
Protein B construct: TRKRV(pSer)LPQW (crude synthetic peptide)
Average holdup BI: 0.2
Holdup BI standard deviation: 0.82
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.13
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 18
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 1457-1466
Protein B site: 14-3-3_pSer1462
Protein B modification: (SEP)1462
Protein B sequence: YQKQQSLPSL
Protein B construct: YQKQQ(pSer)LPSL (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.12
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 19
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 949-958
Protein B site: 14-3-3_pSer954
Protein B modification: (SEP)954
Protein B sequence: GAPVASRSWR
Protein B construct: GAPVA(pSer)RSWR (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 143
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: 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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SHROOM3
Protein B fragment: 1016-1025
Protein B site: 14-3-3_pSer1021
Protein B modification: (SEP)1021
Protein B sequence: KKRSYSEPEK
Protein B construct: KKRSY(pSer)EPEK (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.05
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.