Details of experiments between P31946 and Q9UJF2
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: RASAL2
Protein B fragment: 129-138
Protein B site: 14-3-3_pSer134
Protein B modification: (SEP)134
Protein B sequence: LRRTVSVPSE
Protein B construct: LRRTV(pSer)VPSE (crude synthetic peptide)
Average holdup BI: 0.86
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): 9.58
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: 4.66
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: RASAL2
Protein B fragment: 612-621
Protein B site: 14-3-3_pThr617
Protein B modification: (TPO)617
Protein B sequence: ITKSLTNPTP
Protein B construct: ITKSL(pThr)NPTP (crude synthetic peptide)
Average holdup BI: 0.74
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): 7.8
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: 4.32
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: RASAL2
Protein B fragment: 129-138
Protein B site: 14-3-3_pSer134
Protein B modification: (SEP)134
Protein B sequence: LRRTVSVPSE
Protein B construct: LRRTV(pSer)VPSE (crude synthetic peptide)
Average holdup BI: 0.86
Holdup BI standard deviation: 0.01
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): 7.69
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: 4.79
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: RASAL2
Protein B fragment: 612-621
Protein B site: 14-3-3_pThr617
Protein B modification: (TPO)617
Protein B sequence: ITKSLTNPTP
Protein B construct: ITKSL(pThr)NPTP (crude synthetic peptide)
Average holdup BI: 0.72
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): 7.67
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
Measured pKd: 4.4
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: RASAL2
Protein B fragment: 844-853
Protein B site: 14-3-3_pThr849
Protein B modification: (TPO)849
Protein B sequence: FSRRHTVPDR
Protein B construct: FSRRH(pThr)VPDR (crude synthetic peptide)
Average holdup BI: 0.89
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): 7.19
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
Measured pKd: 4.77
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: RASAL2
Protein B fragment: 844-853
Protein B site: 14-3-3_pThr849
Protein B modification: (TPO)849
Protein B sequence: FSRRHTVPDR
Protein B construct: FSRRH(pThr)VPDR (crude synthetic peptide)
Average holdup BI: 0.88
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): 3.89
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: 4.84
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: RASAL2
Protein B fragment: 882-891
Protein B site: 14-3-3_pSer887
Protein B modification: (SEP)887
Protein B sequence: AKAPPSLPHS
Protein B construct: AKAPP(pSer)LPHS (crude synthetic peptide)
Average holdup BI: 0.15
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): 2.99
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: RASAL2
Protein B fragment: 836-845
Protein B site: 14-3-3_pSer841
Protein B modification: (SEP)841
Protein B sequence: KRSTQSEDFS
Protein B construct: KRSTQ(pSer)EDFS (crude synthetic peptide)
Average holdup BI: 0.38
Holdup BI standard deviation: 0.17
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): 1.84
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: 3.78
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: RASAL2
Protein B fragment: 84-93
Protein B site: 14-3-3_pSer89
Protein B modification: (SEP)89
Protein B sequence: LKESRSHESL
Protein B construct: LKESR(pSer)HESL (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.12
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): 1.76
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: RASAL2
Protein B fragment: 84-93
Protein B site: 14-3-3_pSer89
Protein B modification: (SEP)89
Protein B sequence: LKESRSHESL
Protein B construct: LKESR(pSer)HESL (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.13
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): 1.32
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 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: RASAL2
Protein B fragment: 936-945
Protein B site: 14-3-3_pThr941
Protein B modification: (TPO)941
Protein B sequence: IQRQQTQQVQ
Protein B construct: IQRQQ(pThr)QQVQ (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.23
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): 0.84
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: RASAL2
Protein B fragment: 734-743
Protein B site: 14-3-3_pSer739
Protein B modification: (SEP)739
Protein B sequence: TGSQLSITQV
Protein B construct: TGSQL(pSer)ITQV (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.07
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): 0.62
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
The affinity was below the detection threshold of the assay.
Experiment series 13
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: RASAL2
Protein B fragment: 882-891
Protein B site: 14-3-3_pSer887
Protein B modification: (SEP)887
Protein B sequence: AKAPPSLPHS
Protein B construct: AKAPP(pSer)LPHS (crude synthetic peptide)
Average holdup BI: 0.09
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): 0.43
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: 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: RASAL2
Protein B fragment: 734-743
Protein B site: 14-3-3_pSer739
Protein B modification: (SEP)739
Protein B sequence: TGSQLSITQV
Protein B construct: TGSQL(pSer)ITQV (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.21
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): 0.29
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 15
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: RASAL2
Protein B fragment: 731-740
Protein B site: 14-3-3_pSer736
Protein B modification: (SEP)736
Protein B sequence: IRLTGSQLSI
Protein B construct: IRLTG(pSer)QLSI (crude synthetic peptide)
Average holdup BI: 0.05
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): 0.2
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: 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: RASAL2
Protein B fragment: 731-740
Protein B site: 14-3-3_pSer736
Protein B modification: (SEP)736
Protein B sequence: IRLTGSQLSI
Protein B construct: IRLTG(pSer)QLSI (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.45
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): 0.15
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 17
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: RASAL2
Protein B fragment: 936-945
Protein B site: 14-3-3_pThr941
Protein B modification: (TPO)941
Protein B sequence: IQRQQTQQVQ
Protein B construct: IQRQQ(pThr)QQVQ (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.16
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): 0.09
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 18
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: RASAL2
Protein B fragment: 695-704
Protein B site: 14-3-3_pSer700
Protein B modification: (SEP)700
Protein B sequence: DERESSLPNG
Protein B construct: DERES(pSer)LPNG (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.41
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): 0.04
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.