Details of experiments between Q02241 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: KIF23
Protein A fragment: 600-609
Protein A site: 14-3-3_pSer605
Protein A modification: (SEP)605
Protein A sequence: LQRQFSDKRR
Protein A construct: LQRQF(pSer)DKRR (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): 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 2

Protein A protein: KIF23
Protein A fragment: 938-947
Protein A site: 14-3-3_pSer943
Protein A modification: (SEP)943
Protein A sequence: EMRAGSQLGP
Protein A construct: EMRAG(pSer)QLGP (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): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.65
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 3

Protein A protein: KIF23
Protein A fragment: 155-164
Protein A site: 14-3-3_pSer160
Protein A modification: (SEP)160
Protein A sequence: SNDRNSMDIQ
Protein A construct: SNDRN(pSer)MDIQ (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: 0.25
Holdup BI standard deviation: 0.16
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): 1.47
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: 3.47


Experiment series 4

Protein A protein: KIF23
Protein A fragment: 938-947
Protein A site: 14-3-3_pSer943
Protein A modification: (SEP)943
Protein A sequence: EMRAGSQLGP
Protein A construct: EMRAG(pSer)QLGP (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: 0.17
Holdup BI standard deviation: 0.16
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): 0.98
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 5

Protein A protein: KIF23
Protein A fragment: 13-22
Protein A site: 14-3-3_pSer18
Protein A modification: (SEP)18
Protein A sequence: TVKKGSQTNL
Protein A construct: TVKKG(pSer)QTNL (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: -0.26
Holdup BI standard deviation: 0.41
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): 0.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 6

Protein A protein: KIF23
Protein A fragment: 677-686
Protein A site: 14-3-3_pSer682
Protein A modification: (SEP)682
Protein A sequence: KVTQRSVSPS
Protein A construct: KVTQR(pSer)VSPS (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: 0.19
Holdup BI standard deviation: 0.23
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): 0.49
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 7

Protein A protein: KIF23
Protein A fragment: 155-164
Protein A site: 14-3-3_pSer160
Protein A modification: (SEP)160
Protein A sequence: SNDRNSMDIQ
Protein A construct: SNDRN(pSer)MDIQ (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: 0.54
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): 0.21
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: KIF23
Protein A fragment: 892-901
Protein A site: 14-3-3_pThr897
Protein A modification: (TPO)897
Protein A sequence: FTDIETLKQE
Protein A construct: FTDIE(pThr)LKQE (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: 0.04
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): 0.21
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 9

Protein A protein: KIF23
Protein A fragment: 892-901
Protein A site: 14-3-3_pThr897
Protein A modification: (TPO)897
Protein A sequence: FTDIETLKQE
Protein A construct: FTDIE(pThr)LKQE (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): 0.21
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: KIF23
Protein A fragment: 892-901
Protein A site: 14-3-3_pThr897
Protein A modification: (TPO)897
Protein A sequence: FTDIETLKQE
Protein A construct: FTDIE(pThr)LKQE (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: -0.02
Holdup BI standard deviation: 0.08
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): 0.17
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 11

Protein A protein: KIF23
Protein A fragment: 155-164
Protein A site: 14-3-3_pSer160
Protein A modification: (SEP)160
Protein A sequence: SNDRNSMDIQ
Protein A construct: SNDRN(pSer)MDIQ (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: 0.08
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): 0.17
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 12

Protein A protein: KIF23
Protein A fragment: 922-931
Protein A site: 14-3-3_pThr927
Protein A modification: (TPO)927
Protein A sequence: AESEWTDVET
Protein A construct: AESEW(pThr)DVET (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: -0.13
Holdup BI standard deviation: 0.68
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 13

Protein A protein: KIF23
Protein A fragment: 13-22
Protein A site: 14-3-3_pSer18
Protein A modification: (SEP)18
Protein A sequence: TVKKGSQTNL
Protein A construct: TVKKG(pSer)QTNL (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: 0.85
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): 0.14
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 14

Protein A protein: KIF23
Protein A fragment: 677-686
Protein A site: 14-3-3_pSer682
Protein A modification: (SEP)682
Protein A sequence: KVTQRSVSPS
Protein A construct: KVTQR(pSer)VSPS (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: 0.11
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): 0.11
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: KIF23
Protein A fragment: 600-609
Protein A site: 14-3-3_pSer605
Protein A modification: (SEP)605
Protein A sequence: LQRQFSDKRR
Protein A construct: LQRQF(pSer)DKRR (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: 0.07
Holdup BI standard deviation: 0.55
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 16

Protein A protein: KIF23
Protein A fragment: 13-22
Protein A site: 14-3-3_pSer18
Protein A modification: (SEP)18
Protein A sequence: TVKKGSQTNL
Protein A construct: TVKKG(pSer)QTNL (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: 0.15
Holdup BI standard deviation: 0.85
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): 0.07
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 17

Protein A protein: KIF23
Protein A fragment: 677-686
Protein A site: 14-3-3_pSer682
Protein A modification: (SEP)682
Protein A sequence: KVTQRSVSPS
Protein A construct: KVTQR(pSer)VSPS (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: 0.37
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): 0.03
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 18

Protein A protein: KIF23
Protein A fragment: 938-947
Protein A site: 14-3-3_pSer943
Protein A modification: (SEP)943
Protein A sequence: EMRAGSQLGP
Protein A construct: EMRAG(pSer)QLGP (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: 0.43
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
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.