Details of experiments between P61981 and Q9Y3M2
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CBY1
Protein B fragment: 15-24
Protein B site: 14-3-3_pSer20
Protein B modification: (SEP)20
Protein B sequence: PRKSASLSNL
Protein B construct: PRKSA(pSer)LSNL (crude synthetic peptide)
Average holdup BI: 0.85
Holdup BI standard deviation: 0.01
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): 8.51
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: 4.72
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: CBY1
Protein B fragment: 15-24
Protein B site: 14-3-3_pSer20
Protein B modification: (SEP)20
Protein B sequence: PRKSASLSNL
Protein B construct: PRKSA(pSer)LSNL (crude synthetic peptide)
Average holdup BI: 0.89
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): 8.21
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
Measured pKd: 4.77
Experiment series 3
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: CBY1
Protein B fragment: 15-24
Protein B site: 14-3-3_pSer20
Protein B modification: (SEP)20
Protein B sequence: PRKSASLSNL
Protein B construct: PRKSA(pSer)LSNL (crude synthetic peptide)
Average holdup BI: 0.9
Holdup BI standard deviation: 0.01
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): 7.93
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: 4.84
Experiment series 4
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: CBY1
Protein B fragment: 2-11
Protein B site: 14-3-3_pThr7
Protein B modification: (TPO)7
Protein B sequence: PFFGNTFSPK
Protein B construct: PFFGN(pThr)FSPK (crude synthetic peptide)
Average holdup BI: 0.53
Holdup BI standard deviation: 0.03
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): 6.36
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: 4
Experiment series 5
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: CBY1
Protein B fragment: 2-11
Protein B site: 14-3-3_pThr7
Protein B modification: (TPO)7
Protein B sequence: PFFGNTFSPK
Protein B construct: PFFGN(pThr)FSPK (crude synthetic peptide)
Average holdup BI: 0.63
Holdup BI standard deviation: 0.07
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): 5.6
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: 4.11
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: CBY1
Protein B fragment: 2-11
Protein B site: 14-3-3_pThr7
Protein B modification: (TPO)7
Protein B sequence: PFFGNTFSPK
Protein B construct: PFFGN(pThr)FSPK (crude synthetic peptide)
Average holdup BI: 0.53
Holdup BI standard deviation: 0.09
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): 3.38
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
Measured pKd: 3.93
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: CBY1
Protein B fragment: 21-30
Protein B site: 14-3-3_pSer26
Protein B modification: (SEP)26
Protein B sequence: LSNLHSLDRS
Protein B construct: LSNLH(pSer)LDRS (crude synthetic peptide)
Average holdup BI: -0.09
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): 1.99
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: 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: CBY1
Protein B fragment: 21-30
Protein B site: 14-3-3_pSer26
Protein B modification: (SEP)26
Protein B sequence: LSNLHSLDRS
Protein B construct: LSNLH(pSer)LDRS (crude synthetic peptide)
Average holdup BI: -0.15
Holdup BI standard deviation: 0.15
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): 1.07
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 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: CBY1
Protein B fragment: 21-30
Protein B site: 14-3-3_pSer26
Protein B modification: (SEP)26
Protein B sequence: LSNLHSLDRS
Protein B construct: LSNLH(pSer)LDRS (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.68
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): 0.14
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.