Details of experiments between P61981 and O43379
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: WDR62
Protein B fragment: 1223-1232
Protein B site: 14-3-3_pSer1228
Protein B modification: (SEP)1228
Protein B sequence: ISRSISLGDS
Protein B construct: ISRSI(pSer)LGDS (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): 9.53
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 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: WDR62
Protein B fragment: 1223-1232
Protein B site: 14-3-3_pSer1228
Protein B modification: (SEP)1228
Protein B sequence: ISRSISLGDS
Protein B construct: ISRSI(pSer)LGDS (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): 9.05
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: WDR62
Protein B fragment: 1223-1232
Protein B site: 14-3-3_pSer1228
Protein B modification: (SEP)1228
Protein B sequence: ISRSISLGDS
Protein B construct: ISRSI(pSer)LGDS (crude synthetic peptide)
Average holdup BI: 0.81
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): 7.68
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: 4.51
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: WDR62
Protein B fragment: 1244-1253
Protein B site: 14-3-3_pSer1249
Protein B modification: (SEP)1249
Protein B sequence: LRRPSSVGEL
Protein B construct: LRRPS(pSer)VGEL (crude synthetic peptide)
Average holdup BI: 0.62
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): 6.91
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: 4.17
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: WDR62
Protein B fragment: 1244-1253
Protein B site: 14-3-3_pSer1249
Protein B modification: (SEP)1249
Protein B sequence: LRRPSSVGEL
Protein B construct: LRRPS(pSer)VGEL (crude synthetic peptide)
Average holdup BI: 0.62
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): 6.19
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
Measured pKd: 4.11
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: WDR62
Protein B fragment: 1244-1253
Protein B site: 14-3-3_pSer1249
Protein B modification: (SEP)1249
Protein B sequence: LRRPSSVGEL
Protein B construct: LRRPS(pSer)VGEL (crude synthetic peptide)
Average holdup BI: 0.53
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): 5.63
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: 3.92
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: WDR62
Protein B fragment: 1337-1346
Protein B site: 14-3-3_pSer1342
Protein B modification: (SEP)1342
Protein B sequence: SAFRPSLPAP
Protein B construct: SAFRP(pSer)LPAP (crude synthetic peptide)
Average holdup BI: 0.46
Holdup BI standard deviation: 0.03
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): 5.52
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: 3.88
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: WDR62
Protein B fragment: 1337-1346
Protein B site: 14-3-3_pSer1342
Protein B modification: (SEP)1342
Protein B sequence: SAFRPSLPAP
Protein B construct: SAFRP(pSer)LPAP (crude synthetic peptide)
Average holdup BI: 0.25
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): 3.52
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: WDR62
Protein B fragment: 1337-1346
Protein B site: 14-3-3_pSer1342
Protein B modification: (SEP)1342
Protein B sequence: SAFRPSLPAP
Protein B construct: SAFRP(pSer)LPAP (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.09
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): 1.85
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 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: WDR62
Protein B fragment: 892-901
Protein B site: 14-3-3_pSer897
Protein B modification: (SEP)897
Protein B sequence: ESLENSILDS
Protein B construct: ESLEN(pSer)ILDS (crude synthetic peptide)
Average holdup BI: -0.19
Holdup BI standard deviation: 0.28
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): 0.86
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 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: WDR62
Protein B fragment: 1129-1138
Protein B site: 14-3-3_pSer1134
Protein B modification: (SEP)1134
Protein B sequence: MDRGGSQPRA
Protein B construct: MDRGG(pSer)QPRA (crude synthetic peptide)
Average holdup BI: 0.24
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): 0.74
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: 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: WDR62
Protein B fragment: 892-901
Protein B site: 14-3-3_pSer897
Protein B modification: (SEP)897
Protein B sequence: ESLENSILDS
Protein B construct: ESLEN(pSer)ILDS (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.15
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): 0.45
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 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: WDR62
Protein B fragment: 44-53
Protein B site: 14-3-3_pSer49
Protein B modification: (SEP)49
Protein B sequence: RRTRLSTASE
Protein B construct: RRTRL(pSer)TASE (crude synthetic peptide)
Average holdup BI: -0.17
Holdup BI standard deviation: 0.74
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): 0.18
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 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: WDR62
Protein B fragment: 892-901
Protein B site: 14-3-3_pSer897
Protein B modification: (SEP)897
Protein B sequence: ESLENSILDS
Protein B construct: ESLEN(pSer)ILDS (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.25
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): 0.14
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.