Details of experiments between Q04917 and Q02241
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: KIF23
Protein B fragment: 938-947
Protein B site: 14-3-3_pSer943
Protein B modification: (SEP)943
Protein B sequence: EMRAGSQLGP
Protein B construct: EMRAG(pSer)QLGP (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.81
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 2
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: KIF23
Protein B fragment: 13-22
Protein B site: 14-3-3_pSer18
Protein B modification: (SEP)18
Protein B sequence: TVKKGSQTNL
Protein B construct: TVKKG(pSer)QTNL (crude synthetic peptide)
Average holdup BI: -0.39
Holdup BI standard deviation: 0.5
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.94
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 3
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: KIF23
Protein B fragment: 892-901
Protein B site: 14-3-3_pThr897
Protein B modification: (TPO)897
Protein B sequence: FTDIETLKQE
Protein B construct: FTDIE(pThr)LKQE (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.72
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: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: KIF23
Protein B fragment: 155-164
Protein B site: 14-3-3_pSer160
Protein B modification: (SEP)160
Protein B sequence: SNDRNSMDIQ
Protein B construct: SNDRN(pSer)MDIQ (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.17
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 5
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: KIF23
Protein B fragment: 922-931
Protein B site: 14-3-3_pThr927
Protein B modification: (TPO)927
Protein B sequence: AESEWTDVET
Protein B construct: AESEW(pThr)DVET (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.69
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.02
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.