Details of experiments between P61981 and Q02241
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 892-901
Protein B site: 14-3-3_pThr897
Protein B modification: (TPO)897
Protein B sequence: FTDIETLKQE
Protein B construct: FTDIE(pThr)LKQE (crude synthetic peptide)

Average holdup BI: -0.03
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.83
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 938-947
Protein B site: 14-3-3_pSer943
Protein B modification: (SEP)943
Protein B sequence: EMRAGSQLGP
Protein B construct: EMRAG(pSer)QLGP (crude synthetic peptide)

Average holdup BI: 0.07
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.4
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 938-947
Protein B site: 14-3-3_pSer943
Protein B modification: (SEP)943
Protein B sequence: EMRAGSQLGP
Protein B construct: EMRAG(pSer)QLGP (crude synthetic peptide)

Average holdup BI: 0.23
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.4
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 4

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 155-164
Protein B site: 14-3-3_pSer160
Protein B modification: (SEP)160
Protein B sequence: SNDRNSMDIQ
Protein B construct: SNDRN(pSer)MDIQ (crude synthetic peptide)

Average holdup BI: -0.08
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.09
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 5

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 155-164
Protein B site: 14-3-3_pSer160
Protein B modification: (SEP)160
Protein B sequence: SNDRNSMDIQ
Protein B construct: SNDRN(pSer)MDIQ (crude synthetic peptide)

Average holdup BI: 0.22
Holdup BI standard deviation: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.94
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
The affinity was below the detection threshold of the assay.


Experiment series 6

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 892-901
Protein B site: 14-3-3_pThr897
Protein B modification: (TPO)897
Protein B sequence: FTDIETLKQE
Protein B construct: FTDIE(pThr)LKQE (crude synthetic peptide)

Average holdup BI: -0.04
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.61
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 7

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 862-871
Protein B site: 14-3-3_pSer867
Protein B modification: (SEP)867
Protein B sequence: HQELASDGEI
Protein B construct: HQELA(pSer)DGEI (crude synthetic peptide)

Average holdup BI: 0.23
Holdup BI standard deviation: 0.88
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.14
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 8

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 677-686
Protein B site: 14-3-3_pSer682
Protein B modification: (SEP)682
Protein B sequence: KVTQRSVSPS
Protein B construct: KVTQR(pSer)VSPS (crude synthetic peptide)

Average holdup BI: 0.09
Holdup BI standard deviation: 0.63
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.1
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 9

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 13-22
Protein B site: 14-3-3_pSer18
Protein B modification: (SEP)18
Protein B sequence: TVKKGSQTNL
Protein B construct: TVKKG(pSer)QTNL (crude synthetic peptide)

Average holdup BI: -0.17
Holdup BI standard deviation: 1.18
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 10

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 155-164
Protein B site: 14-3-3_pSer160
Protein B modification: (SEP)160
Protein B sequence: SNDRNSMDIQ
Protein B construct: SNDRN(pSer)MDIQ (crude synthetic peptide)

Average holdup BI: -0.08
Holdup BI standard deviation: 0.72
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.08
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 938-947
Protein B site: 14-3-3_pSer943
Protein B modification: (SEP)943
Protein B sequence: EMRAGSQLGP
Protein B construct: EMRAG(pSer)QLGP (crude synthetic peptide)

Average holdup BI: 0.05
Holdup BI standard deviation: 0.45
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.08
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 892-901
Protein B site: 14-3-3_pThr897
Protein B modification: (TPO)897
Protein B sequence: FTDIETLKQE
Protein B construct: FTDIE(pThr)LKQE (crude synthetic peptide)

Average holdup BI: -0.01
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.05
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 13

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 922-931
Protein B site: 14-3-3_pThr927
Protein B modification: (TPO)927
Protein B sequence: AESEWTDVET
Protein B construct: AESEW(pThr)DVET (crude synthetic peptide)

Average holdup BI: 0.04
Holdup BI standard deviation: 0.64
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.04
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 14

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 862-871
Protein B site: 14-3-3_pSer867
Protein B modification: (SEP)867
Protein B sequence: HQELASDGEI
Protein B construct: HQELA(pSer)DGEI (crude synthetic peptide)

Average holdup BI: -0.14
Holdup BI standard deviation: 2.43
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.04
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 15

Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: KIF23
Protein B fragment: 919-928
Protein B site: 14-3-3_pSer924
Protein B modification: (SEP)924
Protein B sequence: PDGAESEWTD
Protein B construct: PDGAE(pSer)EWTD (crude synthetic peptide)

Average holdup BI: -0.02
Holdup BI standard deviation: 1.1
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.01
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.