Details of experiments between P30305 and P31947
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: CDC25B
Protein A fragment: 225-234
Protein A site: 14-3-3_pSer230
Protein A modification: (SEP)230
Protein A sequence: AQRPSSAPDL
Protein A construct: AQRPS(pSer)APDL (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.69
Holdup BI standard deviation: 0.01
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): 10.54
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: 4.3


Experiment series 2

Protein A protein: CDC25B
Protein A fragment: 225-234
Protein A site: 14-3-3_pSer230
Protein A modification: (SEP)230
Protein A sequence: AQRPSSAPDL
Protein A construct: AQRPS(pSer)APDL (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.8
Holdup BI standard deviation: 0.02
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): 10.48
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: 4.45


Experiment series 3

Protein A protein: CDC25B
Protein A fragment: 146-155
Protein A site: 14-3-3_pSer151
Protein A modification: (SEP)151
Protein A sequence: IRRFQSMPVR
Protein A construct: IRRFQ(pSer)MPVR (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.91
Holdup BI standard deviation: 0.02
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): 8.1
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 4

Protein A protein: CDC25B
Protein A fragment: 146-155
Protein A site: 14-3-3_pSer151
Protein A modification: (SEP)151
Protein A sequence: IRRFQSMPDG
Protein A construct: IRRFQ(pSer)MPDG (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.7
Holdup BI standard deviation: 0.02
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): 7.71
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: 4.2


Experiment series 5

Protein A protein: CDC25B
Protein A fragment: 558-567
Protein A site: 14-3-3_pSer563
Protein A modification: (SEP)563
Protein A sequence: RLKTRSWAGE
Protein A construct: RLKTR(pSer)WAGE (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.72
Holdup BI standard deviation: 0.03
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): 7.43
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
Measured pKd: 4.25


Experiment series 6

Protein A protein: CDC25B
Protein A fragment: 558-567
Protein A site: 14-3-3_pSer563
Protein A modification: (SEP)563
Protein A sequence: RLKTRSWAGE
Protein A construct: RLKTR(pSer)WAGE (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.63
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): 7.05
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: 4.17


Experiment series 7

Protein A protein: CDC25B
Protein A fragment: 225-234
Protein A site: 14-3-3_pSer230
Protein A modification: (SEP)230
Protein A sequence: AQRPSSAPDL
Protein A construct: AQRPS(pSer)APDL (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.75
Holdup BI standard deviation: 0.02
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): 6.51
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
Measured pKd: 4.41


Experiment series 8

Protein A protein: CDC25B
Protein A fragment: 146-155
Protein A site: 14-3-3_pSer151
Protein A modification: (SEP)151
Protein A sequence: IRRFQSMPDG
Protein A construct: IRRFQ(pSer)MPDG (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.65
Holdup BI standard deviation: 0.06
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): 4.94
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: 4.21


Experiment series 9

Protein A protein: CDC25B
Protein A fragment: 146-155
Protein A site: 14-3-3_pSer151
Protein A modification: (SEP)151
Protein A sequence: IRRFQSMPVR
Protein A construct: IRRFQ(pSer)MPVR (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.79
Holdup BI standard deviation: 0.06
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): 4.67
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: 4.53


Experiment series 10

Protein A protein: CDC25B
Protein A fragment: 558-567
Protein A site: 14-3-3_pSer563
Protein A modification: (SEP)563
Protein A sequence: RLKTRSWAGE
Protein A construct: RLKTR(pSer)WAGE (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.62
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): 4.22
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 11

Protein A protein: CDC25B
Protein A fragment: 146-155
Protein A site: 14-3-3_pSer151
Protein A modification: (SEP)151
Protein A sequence: IRRFQSMPVR
Protein A construct: IRRFQ(pSer)MPVR (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.97
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): 3.28
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: 5.44


Experiment series 12

Protein A protein: CDC25B
Protein A fragment: 286-295
Protein A site: 14-3-3_pSer291
Protein A modification: (SEP)291
Protein A sequence: PPGMESLISA
Protein A construct: PPGME(pSer)LISA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.16
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): 2.99
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: CDC25B
Protein A fragment: 53-62
Protein A site: 14-3-3_pThr58
Protein A modification: (TPO)58
Protein A sequence: TTLTQTMHDL
Protein A construct: TTLTQ(pThr)MHDL (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

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): 2.58
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: CDC25B
Protein A fragment: 48-57
Protein A site: 14-3-3_pThr53
Protein A modification: (TPO)53
Protein A sequence: ASSPVTTLTQ
Protein A construct: ASSPV(pThr)TLTQ (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

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): 2.26
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 15

Protein A protein: CDC25B
Protein A fragment: 122-131
Protein A site: 14-3-3_pThr127
Protein A modification: (TPO)127
Protein A sequence: HMAEQTFEQA
Protein A construct: HMAEQ(pThr)FEQA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.21
Holdup BI standard deviation: 0.12
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): 1.73
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: 3.37


Experiment series 16

Protein A protein: CDC25B
Protein A fragment: 460-469
Protein A site: 14-3-3_pSer465
Protein A modification: (SEP)465
Protein A sequence: ERDAESFLLK
Protein A construct: ERDAE(pSer)FLLK (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.07
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 17

Protein A protein: CDC25B
Protein A fragment: 48-57
Protein A site: 14-3-3_pThr53
Protein A modification: (TPO)53
Protein A sequence: ASSPVTTLTQ
Protein A construct: ASSPV(pThr)TLTQ (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.18
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): 1.29
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 18

Protein A protein: CDC25B
Protein A fragment: 122-131
Protein A site: 14-3-3_pThr127
Protein A modification: (TPO)127
Protein A sequence: HMAEQTFEQA
Protein A construct: HMAEQ(pThr)FEQA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.07
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): 1.04
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
The affinity was below the detection threshold of the assay.


Experiment series 19

Protein A protein: CDC25B
Protein A fragment: 286-295
Protein A site: 14-3-3_pSer291
Protein A modification: (SEP)291
Protein A sequence: PPGMESLISA
Protein A construct: PPGME(pSer)LISA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.15
Holdup BI standard deviation: 0.13
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): 1.02
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 20

Protein A protein: CDC25B
Protein A fragment: 399-408
Protein A site: 14-3-3_pThr404
Protein A modification: (TPO)404
Protein A sequence: AFLLQTVDGK
Protein A construct: AFLLQ(pThr)VDGK (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.06
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): 0.91
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
The affinity was below the detection threshold of the assay.


Experiment series 21

Protein A protein: CDC25B
Protein A fragment: 53-62
Protein A site: 14-3-3_pThr58
Protein A modification: (TPO)58
Protein A sequence: TTLTQTMHDL
Protein A construct: TTLTQ(pThr)MHDL (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.15
Holdup BI standard deviation: 0.17
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): 0.88
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 22

Protein A protein: CDC25B
Protein A fragment: 51-60
Protein A site: 14-3-3_pThr56
Protein A modification: (TPO)56
Protein A sequence: PVTTLTQTMH
Protein A construct: PVTTL(pThr)QTMH (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.19
Holdup BI standard deviation: 0.25
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): 0.8
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
The affinity was below the detection threshold of the assay.


Experiment series 23

Protein A protein: CDC25B
Protein A fragment: 370-379
Protein A site: 14-3-3_pSer375
Protein A modification: (SEP)375
Protein A sequence: VLRSKSLCHD
Protein A construct: VLRSK(pSer)LCHD (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.38
Holdup BI standard deviation: 0.53
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): 0.77
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 24

Protein A protein: CDC25B
Protein A fragment: 51-60
Protein A site: 14-3-3_pThr56
Protein A modification: (TPO)56
Protein A sequence: PVTTLTQTMH
Protein A construct: PVTTL(pThr)QTMH (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.12
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): 0.74
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: CDC25B
Protein A fragment: 51-60
Protein A site: 14-3-3_pThr56
Protein A modification: (TPO)56
Protein A sequence: PVTTLTQTMH
Protein A construct: PVTTL(pThr)QTMH (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.07
Holdup BI standard deviation: 0.11
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): 0.68
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 26

Protein A protein: CDC25B
Protein A fragment: 49-58
Protein A site: 14-3-3_pThr54
Protein A modification: (TPO)54
Protein A sequence: SSPVTTLTQT
Protein A construct: SSPVT(pThr)LTQT (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.17
Holdup BI standard deviation: 0.29
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): 0.63
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
The affinity was below the detection threshold of the assay.


Experiment series 27

Protein A protein: CDC25B
Protein A fragment: 399-408
Protein A site: 14-3-3_pThr404
Protein A modification: (TPO)404
Protein A sequence: AFLLQTVDGK
Protein A construct: AFLLQ(pThr)VDGK (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.04
Holdup BI standard deviation: 0.06
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): 0.61
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 28

Protein A protein: CDC25B
Protein A fragment: 146-155
Protein A site: 14-3-3_pSer151
Protein A modification: (SEP)151
Protein A sequence: IRRFQSMPDG
Protein A construct: IRRFQ(pSer)MPDG (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.06
Holdup BI standard deviation: 0.1
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): 0.52
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
The affinity was below the detection threshold of the assay.


Experiment series 29

Protein A protein: CDC25B
Protein A fragment: 286-295
Protein A site: 14-3-3_pSer291
Protein A modification: (SEP)291
Protein A sequence: PPGMESLISA
Protein A construct: PPGME(pSer)LISA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.14
Holdup BI standard deviation: 0.27
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): 0.48
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: CDC25B
Protein A fragment: 122-131
Protein A site: 14-3-3_pThr127
Protein A modification: (TPO)127
Protein A sequence: HMAEQTFEQA
Protein A construct: HMAEQ(pThr)FEQA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.11
Holdup BI standard deviation: 0.28
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): 0.36
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: CDC25B
Protein A fragment: 162-171
Protein A site: 14-3-3_pThr167
Protein A modification: (TPO)167
Protein A sequence: VLRNITNSQA
Protein A construct: VLRNI(pThr)NSQA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.01
Holdup BI standard deviation: 0.03
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): 0.34
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: CDC25B
Protein A fragment: 204-213
Protein A site: 14-3-3_pSer209
Protein A modification: (SEP)209
Protein A sequence: KPTHPSSTHA
Protein A construct: KPTHP(pSer)STHA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.07
Holdup BI standard deviation: 0.17
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): 0.34
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 33

Protein A protein: CDC25B
Protein A fragment: 204-213
Protein A site: 14-3-3_pSer209
Protein A modification: (SEP)209
Protein A sequence: KPTHPSSTHA
Protein A construct: KPTHP(pSer)STHA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.2
Holdup BI standard deviation: 0.64
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): 0.27
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 34

Protein A protein: CDC25B
Protein A fragment: 399-408
Protein A site: 14-3-3_pThr404
Protein A modification: (TPO)404
Protein A sequence: AFLLQTVDGK
Protein A construct: AFLLQ(pThr)VDGK (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.02
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): 0.26
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 35

Protein A protein: CDC25B
Protein A fragment: 48-57
Protein A site: 14-3-3_pThr53
Protein A modification: (TPO)53
Protein A sequence: ASSPVTTLTQ
Protein A construct: ASSPV(pThr)TLTQ (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.01
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): 0.24
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 36

Protein A protein: CDC25B
Protein A fragment: 49-58
Protein A site: 14-3-3_pThr54
Protein A modification: (TPO)54
Protein A sequence: SSPVTTLTQT
Protein A construct: SSPVT(pThr)LTQT (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.04
Holdup BI standard deviation: 0.14
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): 0.24
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 37

Protein A protein: CDC25B
Protein A fragment: 204-213
Protein A site: 14-3-3_pSer209
Protein A modification: (SEP)209
Protein A sequence: KPTHPSSTHA
Protein A construct: KPTHP(pSer)STHA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.08
Holdup BI standard deviation: 0.27
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): 0.23
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 38

Protein A protein: CDC25B
Protein A fragment: 164-173
Protein A site: 14-3-3_pSer169
Protein A modification: (SEP)169
Protein A sequence: RNITNSQAPD
Protein A construct: RNITN(pSer)QAPD (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: -0.02
Holdup BI standard deviation: 0.1
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): 0.17
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: CDC25B
Protein A fragment: 460-469
Protein A site: 14-3-3_pSer465
Protein A modification: (SEP)465
Protein A sequence: ERDAESFLLK
Protein A construct: ERDAE(pSer)FLLK (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.01
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): 0.12
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: CDC25B
Protein A fragment: 164-173
Protein A site: 14-3-3_pSer169
Protein A modification: (SEP)169
Protein A sequence: RNITNSQAPD
Protein A construct: RNITN(pSer)QAPD (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.01
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): 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 41

Protein A protein: CDC25B
Protein A fragment: 370-379
Protein A site: 14-3-3_pSer375
Protein A modification: (SEP)375
Protein A sequence: VLRSKSLCHD
Protein A construct: VLRSK(pSer)LCHD (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.06
Holdup BI standard deviation: 0.73
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): 0.06
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: CDC25B
Protein A fragment: 49-58
Protein A site: 14-3-3_pThr54
Protein A modification: (TPO)54
Protein A sequence: SSPVTTLTQT
Protein A construct: SSPVT(pThr)LTQT (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.01
Holdup BI standard deviation: 0.1
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): 0.05
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: CDC25B
Protein A fragment: 53-62
Protein A site: 14-3-3_pThr58
Protein A modification: (TPO)58
Protein A sequence: TTLTQTMHDL
Protein A construct: TTLTQ(pThr)MHDL (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0.01
Holdup BI standard deviation: 0.28
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): 0.04
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.


Experiment series 44

Protein A protein: CDC25B
Protein A fragment: 460-469
Protein A site: 14-3-3_pSer465
Protein A modification: (SEP)465
Protein A sequence: ERDAESFLLK
Protein A construct: ERDAE(pSer)FLLK (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0
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): 0.03
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 45

Protein A protein: CDC25B
Protein A fragment: 162-171
Protein A site: 14-3-3_pThr167
Protein A modification: (TPO)167
Protein A sequence: VLRNITNSQA
Protein A construct: VLRNI(pThr)NSQA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0
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): 0.03
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.


Experiment series 46

Protein A protein: CDC25B
Protein A fragment: 162-171
Protein A site: 14-3-3_pThr167
Protein A modification: (TPO)167
Protein A sequence: VLRNITNSQA
Protein A construct: VLRNI(pThr)NSQA (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0
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): 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 47

Protein A protein: CDC25B
Protein A fragment: 164-173
Protein A site: 14-3-3_pSer169
Protein A modification: (SEP)169
Protein A sequence: RNITNSQAPD
Protein A construct: RNITN(pSer)QAPD (crude synthetic peptide)

Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma

Average holdup BI: 0
Holdup BI standard deviation: 0.03
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
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.