Details of experiments between P62258 and Q8WUI4
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: HDAC7
Protein B fragment: 481-490
Protein B site: 14-3-3_pSer486
Protein B modification: (SEP)486
Protein B sequence: LSRAQSSPAA
Protein B construct: LSRAQ(pSer)SPAA (crude synthetic peptide)
Average holdup BI: 0.91
Holdup BI standard deviation: 0.01
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): 10.4
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: 4.85
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: HDAC7
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: LRKTVSEPNL
Protein B construct: LRKTV(pSer)EPNL (crude synthetic peptide)
Average holdup BI: 0.91
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): 8.47
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: 4.93
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: HDAC7
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: LRKTVSEPNL
Protein B construct: LRKTV(pSer)EPNL (crude synthetic peptide)
Average holdup BI: 0.91
Holdup BI standard deviation: 0.01
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): 8.13
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: 4.87
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: HDAC7
Protein B fragment: 481-490
Protein B site: 14-3-3_pSer486
Protein B modification: (SEP)486
Protein B sequence: LSRAQSSPAA
Protein B construct: LSRAQ(pSer)SPAA (crude synthetic peptide)
Average holdup BI: 0.9
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): 7.02
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
Measured pKd: 4.89
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: HDAC7
Protein B fragment: 353-362
Protein B site: 14-3-3_pSer358
Protein B modification: (SEP)358
Protein B sequence: LSRTRSEPLP
Protein B construct: LSRTR(pSer)EPLP (crude synthetic peptide)
Average holdup BI: 0.77
Holdup BI standard deviation: 0.05
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): 5.22
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
Measured pKd: 4.47
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: HDAC7
Protein B fragment: 353-362
Protein B site: 14-3-3_pSer358
Protein B modification: (SEP)358
Protein B sequence: LSRTRSEPLP
Protein B construct: LSRTR(pSer)EPLP (crude synthetic peptide)
Average holdup BI: 0.67
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): 4.6
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: 4.15
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: HDAC7
Protein B fragment: 176-185
Protein B site: 14-3-3_pSer181
Protein B modification: (SEP)181
Protein B sequence: LLRKESAPPS
Protein B construct: LLRKE(pSer)APPS (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.05
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): 2.71
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 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: HDAC7
Protein B fragment: 176-185
Protein B site: 14-3-3_pSer181
Protein B modification: (SEP)181
Protein B sequence: LLRKESAPPS
Protein B construct: LLRKE(pSer)APPS (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.07
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): 2.51
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 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: HDAC7
Protein B fragment: 459-468
Protein B site: 14-3-3_pSer464
Protein B modification: (SEP)464
Protein B sequence: RLPRGSTGDT
Protein B construct: RLPRG(pSer)TGDT (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.18
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): 0.28
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: HDAC7
Protein B fragment: 400-409
Protein B site: 14-3-3_pSer405
Protein B modification: (SEP)405
Protein B sequence: LRQIPSAEDL
Protein B construct: LRQIP(pSer)AEDL (crude synthetic peptide)
Average holdup BI: -0.04
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): 0.26
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: HDAC7
Protein B fragment: 400-409
Protein B site: 14-3-3_pSer405
Protein B modification: (SEP)405
Protein B sequence: LRQIPSAEDL
Protein B construct: LRQIP(pSer)AEDL (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.08
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): 0.25
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: HDAC7
Protein B fragment: 502-511
Protein B site: 14-3-3_pSer507
Protein B modification: (SEP)507
Protein B sequence: QARVLSSSET
Protein B construct: QARVL(pSer)SSET (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.07
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): 0.14
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: 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: HDAC7
Protein B fragment: 502-511
Protein B site: 14-3-3_pSer507
Protein B modification: (SEP)507
Protein B sequence: QARVLSSSET
Protein B construct: QARVL(pSer)SSET (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.04
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): 0.14
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 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: HDAC7
Protein B fragment: 459-468
Protein B site: 14-3-3_pSer464
Protein B modification: (SEP)464
Protein B sequence: RLPRGSTGDT
Protein B construct: RLPRG(pSer)TGDT (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.24
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): 0.03
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.