Details of experiments between P31946 and Q9UQB8
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 335-344
Protein B site: 14-3-3_pThr340
Protein B modification: (TPO)340
Protein B sequence: DSYSNTLPVR
Protein B construct: DSYSN(pThr)LPVR (crude synthetic peptide)
Average holdup BI: 0.75
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): 9.62
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: 4.35
Experiment series 2
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 335-344
Protein B site: 14-3-3_pThr340
Protein B modification: (TPO)340
Protein B sequence: DSYSNTLPVR
Protein B construct: DSYSN(pThr)LPVR (crude synthetic peptide)
Average holdup BI: 0.77
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.72
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
Measured pKd: 4.52
Experiment series 3
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 361-370
Protein B site: 14-3-3_pSer366
Protein B modification: (SEP)366
Protein B sequence: LPRSSSMAAG
Protein B construct: LPRSS(pSer)MAAG (crude synthetic peptide)
Average holdup BI: 0.52
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.96
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: 4.02
Experiment series 4
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 361-370
Protein B site: 14-3-3_pSer366
Protein B modification: (SEP)366
Protein B sequence: LPRSSSMAAG
Protein B construct: LPRSS(pSer)MAAG (crude synthetic peptide)
Average holdup BI: 0.69
Holdup BI standard deviation: 0.05
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): 4.59
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
Measured pKd: 4.23
Experiment series 5
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 355-364
Protein B site: 14-3-3_pThr360
Protein B modification: (TPO)360
Protein B sequence: TTENKTLPRS
Protein B construct: TTENK(pThr)LPRS (crude synthetic peptide)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.04
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: 3.5
Experiment series 6
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 515-524
Protein B site: 14-3-3_pThr520
Protein B modification: (TPO)520
Protein B sequence: GTLVSTV---
Protein B construct: SGSGTLVS(pThr)V (crude synthetic peptide)
Average holdup BI: 0.16
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): 0.9
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 7
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 331-340
Protein B site: 14-3-3_pSer336
Protein B modification: (SEP)336
Protein B sequence: SKLSDSYSNT
Protein B construct: SKLSD(pSer)YSNT (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.06
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): 0.77
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 515-524
Protein B site: 14-3-3_pThr520
Protein B modification: (TPO)520
Protein B sequence: GTLVSTV---
Protein B construct: SGSGTLVS(pThr)V (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.55
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 514-523
Protein B site: 14-3-3_pSer519
Protein B modification: (SEP)519
Protein B sequence: SGTLVSTV--
Protein B construct: SGSGTLV(pSer)TV (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.18
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.55
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 10
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 514-523
Protein B site: 14-3-3_pSer519
Protein B modification: (SEP)519
Protein B sequence: SGTLVSTV--
Protein B construct: SGSGTLV(pSer)TV (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.24
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): 0.51
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 331-340
Protein B site: 14-3-3_pSer336
Protein B modification: (SEP)336
Protein B sequence: SKLSDSYSNT
Protein B construct: SKLSD(pSer)YSNT (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.19
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: BAIAP2
Protein B fragment: 355-364
Protein B site: 14-3-3_pThr360
Protein B modification: (TPO)360
Protein B sequence: TTENKTLPRS
Protein B construct: TTENK(pThr)LPRS (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.5
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.