Details of experiments between P31947 and Q92625
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: ANKS1A
Protein B fragment: 621-630
Protein B site: 14-3-3_pSer626
Protein B modification: (SEP)626
Protein B sequence: RSLSKSDSDL
Protein B construct: RSLSK(pSer)DSDL (crude synthetic peptide)

Average holdup BI: 0.68
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): 7.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.28


Experiment series 2

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: ANKS1A
Protein B fragment: 621-630
Protein B site: 14-3-3_pSer626
Protein B modification: (SEP)626
Protein B sequence: RSLSKSDSDL
Protein B construct: RSLSK(pSer)DSDL (crude synthetic peptide)

Average holdup BI: 0.73
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): 6.85
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: 4.39


Experiment series 3

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: ANKS1A
Protein B fragment: 621-630
Protein B site: 14-3-3_pSer626
Protein B modification: (SEP)626
Protein B sequence: RSLSKSDSDL
Protein B construct: RSLSK(pSer)DSDL (crude synthetic peptide)

Average holdup BI: 0.69
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): 3.68
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
Measured pKd: 4.2


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: ANKS1A
Protein B fragment: 658-667
Protein B site: 14-3-3_pSer663
Protein B modification: (SEP)663
Protein B sequence: LEKSPSFASE
Protein B construct: LEKSP(pSer)FASE (crude synthetic peptide)

Average holdup BI: 0.03
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): 1.68
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: 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: ANKS1A
Protein B fragment: 658-667
Protein B site: 14-3-3_pSer663
Protein B modification: (SEP)663
Protein B sequence: LEKSPSFASE
Protein B construct: LEKSP(pSer)FASE (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): 1.65
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 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: ANKS1A
Protein B fragment: 672-681
Protein B site: 14-3-3_pSer677
Protein B modification: (SEP)677
Protein B sequence: EKIMSSIGEG
Protein B construct: EKIMS(pSer)IGEG (crude synthetic peptide)

Average holdup BI: 0.16
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): 1.61
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: 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: ANKS1A
Protein B fragment: 331-340
Protein B site: 14-3-3_pSer336
Protein B modification: (SEP)336
Protein B sequence: SISQKSQGDV
Protein B construct: SISQK(pSer)QGDV (crude synthetic peptide)

Average holdup BI: 0.04
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): 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 8

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: ANKS1A
Protein B fragment: 24-33
Protein B site: 14-3-3_pSer29
Protein B modification: (SEP)29
Protein B sequence: SGKRLSSGFG
Protein B construct: SGKRL(pSer)SGFG (crude synthetic peptide)

Average holdup BI: 0.07
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): 1.23
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 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: ANKS1A
Protein B fragment: 24-33
Protein B site: 14-3-3_pSer29
Protein B modification: (SEP)29
Protein B sequence: SGKRLSSGFG
Protein B construct: SGKRL(pSer)SGFG (crude synthetic peptide)

Average holdup BI: 0.03
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): 1.08
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: 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: ANKS1A
Protein B fragment: 672-681
Protein B site: 14-3-3_pSer677
Protein B modification: (SEP)677
Protein B sequence: EKIMSSIGEG
Protein B construct: EKIMS(pSer)IGEG (crude synthetic peptide)

Average holdup BI: 0.02
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): 0.94
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 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: ANKS1A
Protein B fragment: 455-464
Protein B site: 14-3-3_pThr460
Protein B modification: (TPO)460
Protein B sequence: YELLLTAETK
Protein B construct: YELLL(pThr)AETK (crude synthetic peptide)

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): 0.9
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: ANKS1A
Protein B fragment: 658-667
Protein B site: 14-3-3_pSer663
Protein B modification: (SEP)663
Protein B sequence: LEKSPSFASE
Protein B construct: LEKSP(pSer)FASE (crude synthetic peptide)

Average holdup BI: 0.02
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): 0.82
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: 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: ANKS1A
Protein B fragment: 24-33
Protein B site: 14-3-3_pSer29
Protein B modification: (SEP)29
Protein B sequence: SGKRLSSGFG
Protein B construct: SGKRL(pSer)SGFG (crude synthetic peptide)

Average holdup BI: 0.02
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): 0.56
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: 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: ANKS1A
Protein B fragment: 331-340
Protein B site: 14-3-3_pSer336
Protein B modification: (SEP)336
Protein B sequence: SISQKSQGDV
Protein B construct: SISQK(pSer)QGDV (crude synthetic peptide)

Average holdup BI: -0.03
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): 0.35
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: 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: ANKS1A
Protein B fragment: 672-681
Protein B site: 14-3-3_pSer677
Protein B modification: (SEP)677
Protein B sequence: EKIMSSIGEG
Protein B construct: EKIMS(pSer)IGEG (crude synthetic peptide)

Average holdup BI: -0.06
Holdup BI standard deviation: 0.21
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): 0.22
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
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: ANKS1A
Protein B fragment: 840-849
Protein B site: 14-3-3_pSer845
Protein B modification: (SEP)845
Protein B sequence: -PPRFSQLRC
Protein B construct: PPRF(pSer)QLRCQ (crude synthetic peptide)

Average holdup BI: -0.12
Holdup BI standard deviation: 0.78
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): 0.12
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 17

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: ANKS1A
Protein B fragment: 455-464
Protein B site: 14-3-3_pThr460
Protein B modification: (TPO)460
Protein B sequence: YELLLTAETK
Protein B construct: YELLL(pThr)AETK (crude synthetic peptide)

Average holdup BI: 0.01
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): 0.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 18

Protein A protein: 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: ANKS1A
Protein B fragment: 331-340
Protein B site: 14-3-3_pSer336
Protein B modification: (SEP)336
Protein B sequence: SISQKSQGDV
Protein B construct: SISQK(pSer)QGDV (crude synthetic peptide)

Average holdup BI: -0.01
Holdup BI standard deviation: 0.15
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 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: ANKS1A
Protein B fragment: 840-849
Protein B site: 14-3-3_pSer845
Protein B modification: (SEP)845
Protein B sequence: -PPRFSQLRC
Protein B construct: PPRF(pSer)QLRCQ (crude synthetic peptide)

Average holdup BI: 0.02
Holdup BI standard deviation: 0.8
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.