Details of experiments between P61981 and Q9UJ41
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: RABGEF1
Protein B fragment: 384-393
Protein B site: 14-3-3_pThr389
Protein B modification: (TPO)389
Protein B sequence: YMSGQTSPRK
Protein B construct: YMSGQ(pThr)SPRK (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 115
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: 1
Measured pKd: 4
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: RABGEF1
Protein B fragment: 101-110
Protein B site: 14-3-3_pSer106
Protein B modification: (SEP)106
Protein B sequence: VKKFFSASSR
Protein B construct: VKKFF(pSer)ASSR (crude synthetic peptide)
Average holdup BI: 0.82
Holdup BI standard deviation: 0.02
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): 9.04
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: 4.54
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: RABGEF1
Protein B fragment: 101-110
Protein B site: 14-3-3_pSer106
Protein B modification: (SEP)106
Protein B sequence: VKKFFSASSR
Protein B construct: VKKFF(pSer)ASSR (crude synthetic peptide)
Average holdup BI: 0.82
Holdup BI standard deviation: 0.02
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): 7.86
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: 4.52
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: RABGEF1
Protein B fragment: 101-110
Protein B site: 14-3-3_pSer106
Protein B modification: (SEP)106
Protein B sequence: VKKFFSASSR
Protein B construct: VKKFF(pSer)ASSR (crude synthetic peptide)
Average holdup BI: 0.78
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): 7.56
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: 4.51
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: RABGEF1
Protein B fragment: 127-136
Protein B site: 14-3-3_pSer132
Protein B modification: (SEP)132
Protein B sequence: INRQTSIETD
Protein B construct: INRQT(pSer)IETD (crude synthetic peptide)
Average holdup BI: -0.07
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): 1.9
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 6
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: RABGEF1
Protein B fragment: 127-136
Protein B site: 14-3-3_pSer132
Protein B modification: (SEP)132
Protein B sequence: INRQTSIETD
Protein B construct: INRQT(pSer)IETD (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.04
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): 1.41
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 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: RABGEF1
Protein B fragment: 127-136
Protein B site: 14-3-3_pSer132
Protein B modification: (SEP)132
Protein B sequence: INRQTSIETD
Protein B construct: INRQT(pSer)IETD (crude synthetic peptide)
Average holdup BI: -0.07
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): 1.3
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 construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: RABGEF1
Protein B fragment: 372-381
Protein B site: 14-3-3_pSer377
Protein B modification: (SEP)377
Protein B sequence: QSLNLSQEDF
Protein B construct: QSLNL(pSer)QEDF (crude synthetic peptide)
Average holdup BI: -0.11
Holdup BI standard deviation: 0.14
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): 0.79
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: RABGEF1
Protein B fragment: 384-393
Protein B site: 14-3-3_pThr389
Protein B modification: (TPO)389
Protein B sequence: YMSGQTSPRK
Protein B construct: YMSGQ(pThr)SPRK (crude synthetic peptide)
Average holdup BI: 0.35
Holdup BI standard deviation: 0.46
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): 0.56
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 construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: RABGEF1
Protein B fragment: 108-117
Protein B site: 14-3-3_pSer113
Protein B modification: (SEP)113
Protein B sequence: SSRVGSKKEI
Protein B construct: SSRVG(pSer)KKEI (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.12
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): 0.52
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: 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: RABGEF1
Protein B fragment: 372-381
Protein B site: 14-3-3_pSer377
Protein B modification: (SEP)377
Protein B sequence: QSLNLSQEDF
Protein B construct: QSLNL(pSer)QEDF (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.17
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): 0.31
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: RABGEF1
Protein B fragment: 372-381
Protein B site: 14-3-3_pSer377
Protein B modification: (SEP)377
Protein B sequence: QSLNLSQEDF
Protein B construct: QSLNL(pSer)QEDF (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.09
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): 0.05
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.