Details of experiments between P61981 and O60282
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: KIF5C
Protein B fragment: 914-923
Protein B site: 14-3-3_pSer919
Protein B modification: (SEP)919
Protein B sequence: ARRAHSAQIA
Protein B construct: ARRAH(pSer)AQIA (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 construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIF5C
Protein B fragment: 929-938
Protein B site: 14-3-3_pSer934
Protein B modification: (SEP)934
Protein B sequence: GHYPASSPTA
Protein B construct: GHYPA(pSer)SPTA (crude synthetic peptide)
Average holdup BI: 0.81
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): 7.35
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
Measured pKd: 4.52
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: KIF5C
Protein B fragment: 929-938
Protein B site: 14-3-3_pSer934
Protein B modification: (SEP)934
Protein B sequence: GHYPASSPTA
Protein B construct: GHYPA(pSer)SPTA (crude synthetic peptide)
Average holdup BI: 0.91
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): 5.65
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: 4.85
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: KIF5C
Protein B fragment: 929-938
Protein B site: 14-3-3_pSer934
Protein B modification: (SEP)934
Protein B sequence: GHYPASSPTA
Protein B construct: GHYPA(pSer)SPTA (crude synthetic peptide)
Average holdup BI: 0.77
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): 4.39
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
Measured pKd: 4.48
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: KIF5C
Protein B fragment: 312-321
Protein B site: 14-3-3_pThr317
Protein B modification: (TPO)317
Protein B sequence: AETKSTLMFG
Protein B construct: AETKS(pThr)LMFG (crude synthetic peptide)
Average holdup BI: 0.07
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): 2.09
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 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: KIF5C
Protein B fragment: 591-600
Protein B site: 14-3-3_pSer596
Protein B modification: (SEP)596
Protein B sequence: ISKMKSEVKS
Protein B construct: ISKMK(pSer)EVKS (crude synthetic peptide)
Average holdup BI: 0.07
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): 1.83
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
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: KIF5C
Protein B fragment: 591-600
Protein B site: 14-3-3_pSer596
Protein B modification: (SEP)596
Protein B sequence: ISKMKSEVKS
Protein B construct: ISKMK(pSer)EVKS (crude synthetic peptide)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.12
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): 1.76
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: KIF5C
Protein B fragment: 655-664
Protein B site: 14-3-3_pSer660
Protein B modification: (SEP)660
Protein B sequence: RQLEESQDSL
Protein B construct: RQLEE(pSer)QDSL (crude synthetic peptide)
Average holdup BI: -0.09
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): 1.73
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: KIF5C
Protein B fragment: 605-614
Protein B site: 14-3-3_pSer610
Protein B modification: (SEP)610
Protein B sequence: SKQLESAQMD
Protein B construct: SKQLE(pSer)AQMD (crude synthetic peptide)
Average holdup BI: 0.75
Holdup BI standard deviation: 0.17
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): 1.41
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
Measured pKd: 4.34
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: KIF5C
Protein B fragment: 312-321
Protein B site: 14-3-3_pThr317
Protein B modification: (TPO)317
Protein B sequence: AETKSTLMFG
Protein B construct: AETKS(pThr)LMFG (crude synthetic peptide)
Average holdup BI: 0.2
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): 1.39
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 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: KIF5C
Protein B fragment: 830-839
Protein B site: 14-3-3_pSer835
Protein B modification: (SEP)835
Protein B sequence: QKQKISFLEN
Protein B construct: QKQKI(pSer)FLEN (crude synthetic peptide)
Average holdup BI: -0.04
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): 1.19
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: KIF5C
Protein B fragment: 811-820
Protein B site: 14-3-3_pSer816
Protein B modification: (SEP)816
Protein B sequence: TRVKKSVELD
Protein B construct: TRVKK(pSer)VELD (crude synthetic peptide)
Average holdup BI: -0.17
Holdup BI standard deviation: 0.31
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): 0.67
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIF5C
Protein B fragment: 655-664
Protein B site: 14-3-3_pSer660
Protein B modification: (SEP)660
Protein B sequence: RQLEESQDSL
Protein B construct: RQLEE(pSer)QDSL (crude synthetic peptide)
Average holdup BI: -0.02
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): 0.49
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIF5C
Protein B fragment: 811-820
Protein B site: 14-3-3_pSer816
Protein B modification: (SEP)816
Protein B sequence: TRVKKSVELD
Protein B construct: TRVKK(pSer)VELD (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.11
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): 0.33
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: KIF5C
Protein B fragment: 830-839
Protein B site: 14-3-3_pSer835
Protein B modification: (SEP)835
Protein B sequence: QKQKISFLEN
Protein B construct: QKQKI(pSer)FLEN (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.12
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): 0.32
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 16
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: KIF5C
Protein B fragment: 591-600
Protein B site: 14-3-3_pSer596
Protein B modification: (SEP)596
Protein B sequence: ISKMKSEVKS
Protein B construct: ISKMK(pSer)EVKS (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.45
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): 0.27
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 17
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: KIF5C
Protein B fragment: 610-619
Protein B site: 14-3-3_pSer615
Protein B modification: (SEP)615
Protein B sequence: SAQMDSNRKM
Protein B construct: SAQMD(pSer)NRKM (crude synthetic peptide)
Average holdup BI: 0.2
Holdup BI standard deviation: 0.64
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): 0.24
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 18
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: KIF5C
Protein B fragment: 811-820
Protein B site: 14-3-3_pSer816
Protein B modification: (SEP)816
Protein B sequence: TRVKKSVELD
Protein B construct: TRVKK(pSer)VELD (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.21
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): 0.21
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
The affinity was below the detection threshold of the assay.
Experiment series 19
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: KIF5C
Protein B fragment: 312-321
Protein B site: 14-3-3_pThr317
Protein B modification: (TPO)317
Protein B sequence: AETKSTLMFG
Protein B construct: AETKS(pThr)LMFG (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.29
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): 0.16
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 20
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: KIF5C
Protein B fragment: 655-664
Protein B site: 14-3-3_pSer660
Protein B modification: (SEP)660
Protein B sequence: RQLEESQDSL
Protein B construct: RQLEE(pSer)QDSL (crude synthetic peptide)
Average holdup BI: 0
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): 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.
Experiment series 21
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: KIF5C
Protein B fragment: 605-614
Protein B site: 14-3-3_pSer610
Protein B modification: (SEP)610
Protein B sequence: SKQLESAQMD
Protein B construct: SKQLE(pSer)AQMD (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 1.35
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): 0.01
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 22
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: KIF5C
Protein B fragment: 830-839
Protein B site: 14-3-3_pSer835
Protein B modification: (SEP)835
Protein B sequence: QKQKISFLEN
Protein B construct: QKQKI(pSer)FLEN (crude synthetic peptide)
Average holdup BI: 0
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): 0.01
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.