Details of experiments between P61981 and Q86TI0
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: TBC1D1
Protein B fragment: 232-241
Protein B site: 14-3-3_pSer237
Protein B modification: (SEP)237
Protein B sequence: MRKSFSQPGL
Protein B construct: MRKSF(pSer)QPGL (crude synthetic peptide)
Average holdup BI: 0.89
Holdup BI standard deviation: 0.05
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): 4.75
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: 4.85
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: TBC1D1
Protein B fragment: 561-570
Protein B site: 14-3-3_pSer566
Protein B modification: (SEP)566
Protein B sequence: KLLGSSEDLS
Protein B construct: KLLGS(pSer)EDLS (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.01
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): 4.7
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 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: TBC1D1
Protein B fragment: 622-631
Protein B site: 14-3-3_pSer627
Protein B modification: (SEP)627
Protein B sequence: LMRYHSVSTE
Protein B construct: LMRYH(pSer)VSTE (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.08
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): 3.13
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
The affinity was below the detection threshold of the assay.
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: TBC1D1
Protein B fragment: 622-631
Protein B site: 14-3-3_pSer627
Protein B modification: (SEP)627
Protein B sequence: LMRYHSVSTE
Protein B construct: LMRYH(pSer)VSTE (crude synthetic peptide)
Average holdup BI: 0.43
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): 2.97
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: 3.76
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: TBC1D1
Protein B fragment: 232-241
Protein B site: 14-3-3_pSer237
Protein B modification: (SEP)237
Protein B sequence: MRKSFSQPGL
Protein B construct: MRKSF(pSer)QPGL (crude synthetic peptide)
Average holdup BI: 0.9
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): 2.72
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: 4.84
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: TBC1D1
Protein B fragment: 232-241
Protein B site: 14-3-3_pSer237
Protein B modification: (SEP)237
Protein B sequence: MRKSFSQPGL
Protein B construct: MRKSF(pSer)QPGL (crude synthetic peptide)
Average holdup BI: 0.6
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): 1.96
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: 4.07
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: TBC1D1
Protein B fragment: 561-570
Protein B site: 14-3-3_pSer566
Protein B modification: (SEP)566
Protein B sequence: KLLGSSEDLS
Protein B construct: KLLGS(pSer)EDLS (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.04
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): 1.35
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 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: TBC1D1
Protein B fragment: 561-570
Protein B site: 14-3-3_pSer566
Protein B modification: (SEP)566
Protein B sequence: KLLGSSEDLS
Protein B construct: KLLGS(pSer)EDLS (crude synthetic peptide)
Average holdup BI: 0.04
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): 1.01
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
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: TBC1D1
Protein B fragment: 566-575
Protein B site: 14-3-3_pSer571
Protein B modification: (SEP)571
Protein B sequence: SEDLSSDSES
Protein B construct: SEDLS(pSer)DSES (crude synthetic peptide)
Average holdup BI: -0.25
Holdup BI standard deviation: 0.66
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): 0.32
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: TBC1D1
Protein B fragment: 591-600
Protein B site: 14-3-3_pThr596
Protein B modification: (TPO)596
Protein B sequence: RRRANTLSHF
Protein B construct: RRRAN(pThr)LSHF (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.55
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): 0.23
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: TBC1D1
Protein B fragment: 566-575
Protein B site: 14-3-3_pSer571
Protein B modification: (SEP)571
Protein B sequence: SEDLSSDSES
Protein B construct: SEDLS(pSer)DSES (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 1.43
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): 0.05
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.
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: TBC1D1
Protein B fragment: 498-507
Protein B site: 14-3-3_pSer503
Protein B modification: (SEP)503
Protein B sequence: NKAKRSLTES
Protein B construct: NKAKR(pSer)LTES (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 1.06
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): 0.03
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.