Details of experiments between P61981 and Q6P1L5
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: FAM117B
Protein B fragment: 451-460
Protein B site: 14-3-3_pThr456
Protein B modification: (TPO)456
Protein B sequence: LPKYATSPKP
Protein B construct: LPKYA(pThr)SPKP (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration in holdup experiment (10-6M): 134
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: 5
Experiment series 2
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: FAM117B
Protein B fragment: 215-224
Protein B site: 14-3-3_pSer220
Protein B modification: (SEP)220
Protein B sequence: IRRTSSLDTL
Protein B construct: IRRTS(pSer)LDTL (crude synthetic peptide)
Average holdup BI: 0.77
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): 9.1
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: 4.48
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: FAM117B
Protein B fragment: 215-224
Protein B site: 14-3-3_pSer220
Protein B modification: (SEP)220
Protein B sequence: IRRTSSLDTL
Protein B construct: IRRTS(pSer)LDTL (crude synthetic peptide)
Average holdup BI: 0.83
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): 8.8
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: 4.58
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: FAM117B
Protein B fragment: 215-224
Protein B site: 14-3-3_pSer220
Protein B modification: (SEP)220
Protein B sequence: IRRTSSLDTL
Protein B construct: IRRTS(pSer)LDTL (crude synthetic peptide)
Average holdup BI: 0.79
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): 8.65
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: 4.45
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: FAM117B
Protein B fragment: 451-460
Protein B site: 14-3-3_pThr456
Protein B modification: (TPO)456
Protein B sequence: LPKYATSPKP
Protein B construct: LPKYA(pThr)SPKP (crude synthetic peptide)
Average holdup BI: 0.93
Holdup BI standard deviation: 0.02
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): 6.71
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
Measured pKd: 5.08
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: FAM117B
Protein B fragment: 33-42
Protein B site: 14-3-3_pThr38
Protein B modification: (TPO)38
Protein B sequence: QPMRATVPFQ
Protein B construct: QPMRA(pThr)VPFQ (crude synthetic peptide)
Average holdup BI: 0.86
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): 6.36
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: 4.68
Experiment series 7
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: FAM117B
Protein B fragment: 33-42
Protein B site: 14-3-3_pThr38
Protein B modification: (TPO)38
Protein B sequence: QPMRATVPFQ
Protein B construct: QPMRA(pThr)VPFQ (crude synthetic peptide)
Average holdup BI: 0.87
Holdup BI standard deviation: 0.06
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): 5.67
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: 4.78
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: FAM117B
Protein B fragment: 33-42
Protein B site: 14-3-3_pThr38
Protein B modification: (TPO)38
Protein B sequence: QPMRATVPFQ
Protein B construct: QPMRA(pThr)VPFQ (crude synthetic peptide)
Average holdup BI: 0.86
Holdup BI standard deviation: 0.03
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): 4.6
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: 4.68
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: FAM117B
Protein B fragment: 451-460
Protein B site: 14-3-3_pThr456
Protein B modification: (TPO)456
Protein B sequence: LPKYATSPKP
Protein B construct: LPKYA(pThr)SPKP (crude synthetic peptide)
Average holdup BI: 0.96
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): 3.32
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: 5.29
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: FAM117B
Protein B fragment: 204-213
Protein B site: 14-3-3_pSer209
Protein B modification: (SEP)209
Protein B sequence: TRQPSSSPSS
Protein B construct: TRQPS(pSer)SPSS (crude synthetic peptide)
Average holdup BI: -0.37
Holdup BI standard deviation: 0.78
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): 0.59
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: FAM117B
Protein B fragment: 386-395
Protein B site: 14-3-3_pSer391
Protein B modification: (SEP)391
Protein B sequence: SSSTRSIDTQ
Protein B construct: SSSTR(pSer)IDTQ (crude synthetic peptide)
Average holdup BI: -0.23
Holdup BI standard deviation: 0.59
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): 0.39
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 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: FAM117B
Protein B fragment: 386-395
Protein B site: 14-3-3_pSer391
Protein B modification: (SEP)391
Protein B sequence: SSSTRSIDTQ
Protein B construct: SSSTR(pSer)IDTQ (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.4
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): 0.29
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: FAM117B
Protein B fragment: 268-277
Protein B site: 14-3-3_pSer273
Protein B modification: (SEP)273
Protein B sequence: HKRSASWGST
Protein B construct: HKRSA(pSer)WGST (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.72
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): 0.11
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: 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: FAM117B
Protein B fragment: 386-395
Protein B site: 14-3-3_pSer391
Protein B modification: (SEP)391
Protein B sequence: SSSTRSIDTQ
Protein B construct: SSSTR(pSer)IDTQ (crude synthetic peptide)
Average holdup BI: 0.01
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): 0.06
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 15
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: FAM117B
Protein B fragment: 268-277
Protein B site: 14-3-3_pSer273
Protein B modification: (SEP)273
Protein B sequence: HKRSASWGST
Protein B construct: HKRSA(pSer)WGST (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 2.15
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
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.