Details of experiments between P31947 and Q12802
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2406-2415
Protein B site: 14-3-3_pThr2411
Protein B modification: (TPO)2411
Protein B sequence: LFRSNTEEAL
Protein B construct: biotin-ttds-LFRSN(pThr)EEAL (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): below threshold
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022.02.09
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2462-2471
Protein B site: 14-3-3_pThr2467
Protein B modification: (TPO)2467
Protein B sequence: PRRAETFGGF
Protein B construct: PRRAE(pThr)FGGF (crude synthetic peptide)
Average holdup BI: 0.96
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.65
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: 5.33
Experiment series 3
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2462-2471
Protein B site: 14-3-3_pThr2467
Protein B modification: (TPO)2467
Protein B sequence: PRRAETFGGF
Protein B construct: PRRAE(pThr)FGGF (crude synthetic peptide)
Average holdup BI: 0.95
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.46
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: 5.26
Experiment series 4
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2462-2471
Protein B site: 14-3-3_pThr2467
Protein B modification: (TPO)2467
Protein B sequence: PRRAETFGGF
Protein B construct: PRRAE(pThr)FGGF (crude synthetic peptide)
Average holdup BI: 0.95
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.39
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: 5.17
Experiment series 5
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1924-1933
Protein B site: 14-3-3_pSer1929
Protein B modification: (SEP)1929
Protein B sequence: KFLSHSTDSL
Protein B construct: KFLSH(pSer)TDSL (crude synthetic peptide)
Average holdup BI: 0.34
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.19
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: 3.56
Experiment series 6
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2493-2502
Protein B site: 14-3-3_pSer2498
Protein B modification: (SEP)2498
Protein B sequence: LRRTESDSGL
Protein B construct: LRRTE(pSer)DSGL (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.97
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1924-1933
Protein B site: 14-3-3_pSer1929
Protein B modification: (SEP)1929
Protein B sequence: KFLSHSTDSL
Protein B construct: KFLSH(pSer)TDSL (crude synthetic peptide)
Average holdup BI: 0.27
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.76
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: 3.51
Experiment series 8
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1871-1880
Protein B site: 14-3-3_pSer1876
Protein B modification: (SEP)1876
Protein B sequence: KERPRSAVLL
Protein B construct: KERPR(pSer)AVLL (crude synthetic peptide)
Average holdup BI: 0.43
Holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.2
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
Measured pKd: 3.82
Experiment series 9
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1871-1880
Protein B site: 14-3-3_pSer1876
Protein B modification: (SEP)1876
Protein B sequence: KERPRSAVLL
Protein B construct: KERPR(pSer)AVLL (crude synthetic peptide)
Average holdup BI: 0.47
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.04
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: 3.8
Experiment series 10
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2493-2502
Protein B site: 14-3-3_pSer2498
Protein B modification: (SEP)2498
Protein B sequence: LRRTESDSGL
Protein B construct: LRRTE(pSer)DSGL (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.37
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1502-1511
Protein B site: 14-3-3_pSer1507
Protein B modification: (SEP)1507
Protein B sequence: SRDRQSLDGF
Protein B construct: SRDRQ(pSer)LDGF (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.33
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 12
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1502-1511
Protein B site: 14-3-3_pSer1507
Protein B modification: (SEP)1507
Protein B sequence: SRDRQSLDGF
Protein B construct: SRDRQ(pSer)LDGF (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.3
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 13
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1745-1754
Protein B site: 14-3-3_pSer1750
Protein B modification: (SEP)1750
Protein B sequence: VSRTFSYIKN
Protein B construct: VSRTF(pSer)YIKN (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.27
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2493-2502
Protein B site: 14-3-3_pSer2498
Protein B modification: (SEP)2498
Protein B sequence: LRRTESDSGL
Protein B construct: LRRTE(pSer)DSGL (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.25
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2562-2571
Protein B site: 14-3-3_pSer2567
Protein B modification: (SEP)2567
Protein B sequence: LSRPSSLIEQ
Protein B construct: LSRPS(pSer)LIEQ (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.24
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2406-2415
Protein B site: 14-3-3_pThr2411
Protein B modification: (TPO)2411
Protein B sequence: LFRSNTEEAL
Protein B construct: LFRSN(pThr)EEAL (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.95
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1924-1933
Protein B site: 14-3-3_pSer1929
Protein B modification: (SEP)1929
Protein B sequence: KFLSHSTDSL
Protein B construct: KFLSH(pSer)TDSL (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.93
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1221-1230
Protein B site: 14-3-3_pSer1226
Protein B modification: (SEP)1226
Protein B sequence: SGRERSTPSL
Protein B construct: SGRER(pSer)TPSL (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.77
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 19
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2558-2567
Protein B site: 14-3-3_pSer2563
Protein B modification: (SEP)2563
Protein B sequence: LTRSLSRPSS
Protein B construct: LTRSL(pSer)RPSS (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 113
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: 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 20
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1637-1646
Protein B site: 14-3-3_pSer1642
Protein B modification: (SEP)1642
Protein B sequence: EERVDSLVSL
Protein B construct: EERVD(pSer)LVSL (crude synthetic peptide)
Average holdup BI: -0.17
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.67
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
The affinity was below the detection threshold of the assay.
Experiment series 21
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1502-1511
Protein B site: 14-3-3_pSer1507
Protein B modification: (SEP)1507
Protein B sequence: SRDRQSLDGF
Protein B construct: SRDRQ(pSer)LDGF (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.62
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 22
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1871-1880
Protein B site: 14-3-3_pSer1876
Protein B modification: (SEP)1876
Protein B sequence: KERPRSAVLL
Protein B construct: KERPR(pSer)AVLL (crude synthetic peptide)
Average holdup BI: 0.43
Holdup BI standard deviation: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.58
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: 3.83
Experiment series 23
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1221-1230
Protein B site: 14-3-3_pSer1226
Protein B modification: (SEP)1226
Protein B sequence: SGRERSTPSL
Protein B construct: SGRER(pSer)TPSL (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.55
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 24
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1745-1754
Protein B site: 14-3-3_pSer1750
Protein B modification: (SEP)1750
Protein B sequence: VSRTFSYIKN
Protein B construct: VSRTF(pSer)YIKN (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.55
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 25
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1640-1649
Protein B site: 14-3-3_pSer1645
Protein B modification: (SEP)1645
Protein B sequence: VDSLVSLSEE
Protein B construct: VDSLV(pSer)LSEE (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.5
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 26
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2562-2571
Protein B site: 14-3-3_pSer2567
Protein B modification: (SEP)2567
Protein B sequence: LSRPSSLIEQ
Protein B construct: LSRPS(pSer)LIEQ (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.49
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 27
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1640-1649
Protein B site: 14-3-3_pSer1645
Protein B modification: (SEP)1645
Protein B sequence: VDSLVSLSEE
Protein B construct: VDSLV(pSer)LSEE (crude synthetic peptide)
Average holdup BI: -0.24
Holdup BI standard deviation: 0.25
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.47
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 28
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1890-1899
Protein B site: 14-3-3_pSer1895
Protein B modification: (SEP)1895
Protein B sequence: FANRRSQQSV
Protein B construct: FANRR(pSer)QQSV (crude synthetic peptide)
Average holdup BI: -0.18
Holdup BI standard deviation: 0.19
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.1
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 29
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2562-2571
Protein B site: 14-3-3_pSer2567
Protein B modification: (SEP)2567
Protein B sequence: LSRPSSLIEQ
Protein B construct: LSRPS(pSer)LIEQ (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 111
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: 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 30
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1221-1230
Protein B site: 14-3-3_pSer1226
Protein B modification: (SEP)1226
Protein B sequence: SGRERSTPSL
Protein B construct: SGRER(pSer)TPSL (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.83
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 31
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2558-2567
Protein B site: 14-3-3_pSer2563
Protein B modification: (SEP)2563
Protein B sequence: LTRSLSRPSS
Protein B construct: LTRSL(pSer)RPSS (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 142
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 32
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1927-1936
Protein B site: 14-3-3_pSer1932
Protein B modification: (SEP)1932
Protein B sequence: SHSTDSLNKI
Protein B construct: SHSTD(pSer)LNKI (crude synthetic peptide)
Average holdup BI: -0.3
Holdup BI standard deviation: 0.49
Immobilized partner concentration in holdup experiment (10-6M): 111
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: 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 33
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1927-1936
Protein B site: 14-3-3_pSer1932
Protein B modification: (SEP)1932
Protein B sequence: SHSTDSLNKI
Protein B construct: SHSTD(pSer)LNKI (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.64
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 34
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1890-1899
Protein B site: 14-3-3_pSer1895
Protein B modification: (SEP)1895
Protein B sequence: FANRRSQQSV
Protein B construct: FANRR(pSer)QQSV (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.58
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
The affinity was below the detection threshold of the assay.
Experiment series 35
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2406-2415
Protein B site: 14-3-3_pThr2411
Protein B modification: (TPO)2411
Protein B sequence: LFRSNTEEAL
Protein B construct: LFRSN(pThr)EEAL (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 113
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: 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 36
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2406-2415
Protein B site: 14-3-3_pThr2411
Protein B modification: (TPO)2411
Protein B sequence: LFRSNTEEAL
Protein B construct: LFRSN(pThr)EEAL (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 111
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: 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 37
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2468-2477
Protein B site: 14-3-3_pSer2473
Protein B modification: (SEP)2473
Protein B sequence: FGGFDSHQMN
Protein B construct: FGGFD(pSer)HQMN (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.29
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.42
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 38
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2558-2567
Protein B site: 14-3-3_pSer2563
Protein B modification: (SEP)2563
Protein B sequence: LTRSLSRPSS
Protein B construct: LTRSL(pSer)RPSS (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.4
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 39
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1745-1754
Protein B site: 14-3-3_pSer1750
Protein B modification: (SEP)1750
Protein B sequence: VSRTFSYIKN
Protein B construct: VSRTF(pSer)YIKN (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.32
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 40
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1640-1649
Protein B site: 14-3-3_pSer1645
Protein B modification: (SEP)1645
Protein B sequence: VDSLVSLSEE
Protein B construct: VDSLV(pSer)LSEE (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.31
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 41
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2468-2477
Protein B site: 14-3-3_pSer2473
Protein B modification: (SEP)2473
Protein B sequence: FGGFDSHQMN
Protein B construct: FGGFD(pSer)HQMN (crude synthetic peptide)
Average holdup BI: -0.13
Holdup BI standard deviation: 0.37
Immobilized partner concentration in holdup experiment (10-6M): 111
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: 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 42
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1597-1606
Protein B site: 14-3-3_pSer1602
Protein B modification: (SEP)1602
Protein B sequence: HRRSFSLEGL
Protein B construct: HRRSF(pSer)LEGL (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.46
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.28
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 43
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1890-1899
Protein B site: 14-3-3_pSer1895
Protein B modification: (SEP)1895
Protein B sequence: FANRRSQQSV
Protein B construct: FANRR(pSer)QQSV (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 142
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 44
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1637-1646
Protein B site: 14-3-3_pSer1642
Protein B modification: (SEP)1642
Protein B sequence: EERVDSLVSL
Protein B construct: EERVD(pSer)LVSL (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 113
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: 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 45
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1927-1936
Protein B site: 14-3-3_pSer1932
Protein B modification: (SEP)1932
Protein B sequence: SHSTDSLNKI
Protein B construct: SHSTD(pSer)LNKI (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.07
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 46
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 1637-1646
Protein B site: 14-3-3_pSer1642
Protein B modification: (SEP)1642
Protein B sequence: EERVDSLVSL
Protein B construct: EERVD(pSer)LVSL (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.05
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
The affinity was below the detection threshold of the assay.
Experiment series 47
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2340-2349
Protein B site: 14-3-3_pSer2345
Protein B modification: (SEP)2345
Protein B sequence: DEGIPSENEE
Protein B construct: DEGIP(pSer)ENEE (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.86
Immobilized partner concentration in holdup experiment (10-6M): 111
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: 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 48
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: AKAP13
Protein B fragment: 2468-2477
Protein B site: 14-3-3_pSer2473
Protein B modification: (SEP)2473
Protein B sequence: FGGFDSHQMN
Protein B construct: FGGFD(pSer)HQMN (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 142
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.