Details of experiments between P61981 and Q6ICG6
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 284-293
Protein B site: 14-3-3_pSer289
Protein B modification: (SEP)289
Protein B sequence: RVTSFSTPPT
Protein B construct: RVTSF(pSer)TPPT (crude synthetic peptide)
Average holdup BI: 0.95
Holdup BI standard deviation: 0
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): 9.86
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: 5.17
Experiment series 2
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 284-293
Protein B site: 14-3-3_pSer289
Protein B modification: (SEP)289
Protein B sequence: RVTSFSTPPT
Protein B construct: RVTSF(pSer)TPPT (crude synthetic peptide)
Average holdup BI: 0.96
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 9.83
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
Measured pKd: 5.23
Experiment series 3
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 284-293
Protein B site: 14-3-3_pSer289
Protein B modification: (SEP)289
Protein B sequence: RVTSFSTPPT
Protein B construct: RVTSF(pSer)TPPT (crude synthetic peptide)
Average holdup BI: 0.97
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 9.01
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
Measured pKd: 5.39
Experiment series 4
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 301-310
Protein B site: 14-3-3_pSer306
Protein B modification: (SEP)306
Protein B sequence: AFFSPSLKRK
Protein B construct: AFFSP(pSer)LKRK (crude synthetic peptide)
Average holdup BI: 0.22
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.56
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 5
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 301-310
Protein B site: 14-3-3_pSer306
Protein B modification: (SEP)306
Protein B sequence: AFFSPSLKRK
Protein B construct: AFFSP(pSer)LKRK (crude synthetic peptide)
Average holdup BI: 0.37
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.89
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: 3.73
Experiment series 6
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 301-310
Protein B site: 14-3-3_pSer306
Protein B modification: (SEP)306
Protein B sequence: AFFSPSLKRK
Protein B construct: AFFSP(pSer)LKRK (crude synthetic peptide)
Average holdup BI: 0.12
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): 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 7
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 257-266
Protein B site: 14-3-3_pSer262
Protein B modification: (SEP)262
Protein B sequence: AVSRVSTGDT
Protein B construct: AVSRV(pSer)TGDT (crude synthetic peptide)
Average holdup BI: 0.06
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): 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 8
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 257-266
Protein B site: 14-3-3_pSer262
Protein B modification: (SEP)262
Protein B sequence: AVSRVSTGDT
Protein B construct: AVSRV(pSer)TGDT (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.76
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 399-408
Protein B site: 14-3-3_pThr404
Protein B modification: (TPO)404
Protein B sequence: KPILMT----
Protein B construct: EVRQKPILM(pThr) (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.13
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): 0.41
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 257-266
Protein B site: 14-3-3_pSer262
Protein B modification: (SEP)262
Protein B sequence: AVSRVSTGDT
Protein B construct: AVSRV(pSer)TGDT (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 115
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: 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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 399-408
Protein B site: 14-3-3_pThr404
Protein B modification: (TPO)404
Protein B sequence: KPILMT----
Protein B construct: EVRQKPILM(pThr) (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.18
Immobilized partner concentration in holdup experiment (10-6M): 129
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 12
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 357-366
Protein B site: 14-3-3_pSer362
Protein B modification: (SEP)362
Protein B sequence: RSLVGSWLKL
Protein B construct: RSLVG(pSer)WLKL (crude synthetic peptide)
Average holdup BI: -0.22
Holdup BI standard deviation: 1.37
Immobilized partner concentration in holdup experiment (10-6M): 129
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 284-293
Protein B site: 14-3-3_pSer289
Protein B modification: (SEP)289
Protein B sequence: RVTSFSTPPT
Protein B construct: biotin-ttds-RVTSF(pSer)TPPT (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 6.3
Standard deviation of dissociation constant (10-6M): 0.7
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022.01.18
Number of measurements: 3
Measured pKd: 5.2
Experiment series 14
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIAA0930
Protein B fragment: 284-293
Protein B site: 14-3-3_pSer289
Protein B modification: (SEP)289
Protein B sequence: RVTSFSTPPT
Protein B construct: biotin-ttds-RVTSF(pSer)TPPT (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 6.2
Standard deviation of dissociation constant (10-6M): 0.7
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022.01.10
Number of measurements: 3
Measured pKd: 5.21