Details of experiments between P61981 and Q12840
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: KIF5A
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: TKTNLSVHED
Protein B construct: TKTNL(pSer)VHED (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.06
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): 1.01
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.
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: KIF5A
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: TKTNLSVHED
Protein B construct: TKTNL(pSer)VHED (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.1
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): 0.88
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: KIF5A
Protein B fragment: 802-811
Protein B site: 14-3-3_pThr807
Protein B modification: (TPO)807
Protein B sequence: VQDVTTRVKK
Protein B construct: VQDVT(pThr)RVKK (crude synthetic peptide)
Average holdup BI: -0.07
Holdup BI standard deviation: 0.11
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): 0.62
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: KIF5A
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: TKTNLSVHED
Protein B construct: TKTNL(pSer)VHED (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.08
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): 0.23
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 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: KIF5A
Protein B fragment: 802-811
Protein B site: 14-3-3_pThr807
Protein B modification: (TPO)807
Protein B sequence: VQDVTTRVKK
Protein B construct: VQDVT(pThr)RVKK (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.14
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 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: KIF5A
Protein B fragment: 802-811
Protein B site: 14-3-3_pThr807
Protein B modification: (TPO)807
Protein B sequence: VQDVTTRVKK
Protein B construct: VQDVT(pThr)RVKK (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.29
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
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.