Details of experiments between P31947 and Q92974
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: ARHGEF2
Protein B fragment: 881-890
Protein B site: 14-3-3_pSer886
Protein B modification: (SEP)886
Protein B sequence: DPRRRSLPAG
Protein B construct: DPRRR(pSer)LPAG (crude synthetic peptide)

Average holdup BI: 0.43
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): 6.92
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 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: ARHGEF2
Protein B fragment: 881-890
Protein B site: 14-3-3_pSer886
Protein B modification: (SEP)886
Protein B sequence: DPRRRSLPAG
Protein B construct: DPRRR(pSer)LPAG (crude synthetic peptide)

Average holdup BI: 0.49
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): 6.04
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: 3.83


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: ARHGEF2
Protein B fragment: 181-190
Protein B site: 14-3-3_pSer186
Protein B modification: (SEP)186
Protein B sequence: RNRTLSVESL
Protein B construct: RNRTL(pSer)VESL (crude synthetic peptide)

Average holdup BI: 0.33
Holdup BI standard deviation: 0.03
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): 5.84
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: 3.64


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: ARHGEF2
Protein B fragment: 146-155
Protein B site: 14-3-3_pSer151
Protein B modification: (SEP)151
Protein B sequence: LAKSVSTTNI
Protein B construct: LAKSV(pSer)TTNI (crude synthetic peptide)

Average holdup BI: 0.44
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): 5.74
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
Measured pKd: 3.74


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: ARHGEF2
Protein B fragment: 104-113
Protein B site: 14-3-3_pSer109
Protein B modification: (SEP)109
Protein B sequence: ALQSVSLRSK
Protein B construct: ALQSV(pSer)LRSK (crude synthetic peptide)

Average holdup BI: 0.5
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): 5.73
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: 3.84


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: ARHGEF2
Protein B fragment: 146-155
Protein B site: 14-3-3_pSer151
Protein B modification: (SEP)151
Protein B sequence: LAKSVSTTNI
Protein B construct: LAKSV(pSer)TTNI (crude synthetic peptide)

Average holdup BI: 0.33
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): 5.58
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: 3.64


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: ARHGEF2
Protein B fragment: 146-155
Protein B site: 14-3-3_pSer151
Protein B modification: (SEP)151
Protein B sequence: LAKSVSTTNI
Protein B construct: LAKSV(pSer)TTNI (crude synthetic peptide)

Average holdup BI: 0.33
Holdup BI standard deviation: 0.03
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): 5.54
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: 3.65


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: ARHGEF2
Protein B fragment: 117-126
Protein B site: 14-3-3_pSer122
Protein B modification: (SEP)122
Protein B sequence: RERPSSAIYP
Protein B construct: RERPS(pSer)AIYP (crude synthetic peptide)

Average holdup BI: 0.45
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): 5.47
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
Measured pKd: 3.76


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: ARHGEF2
Protein B fragment: 104-113
Protein B site: 14-3-3_pSer109
Protein B modification: (SEP)109
Protein B sequence: ALQSVSLRSK
Protein B construct: ALQSV(pSer)LRSK (crude synthetic peptide)

Average holdup BI: 0.4
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): 5.39
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: 3.76


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: ARHGEF2
Protein B fragment: 104-113
Protein B site: 14-3-3_pSer109
Protein B modification: (SEP)109
Protein B sequence: ALQSVSLRSK
Protein B construct: ALQSV(pSer)LRSK (crude synthetic peptide)

Average holdup BI: 0.37
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): 5.21
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
Measured pKd: 3.72


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: ARHGEF2
Protein B fragment: 181-190
Protein B site: 14-3-3_pSer186
Protein B modification: (SEP)186
Protein B sequence: RNRTLSVESL
Protein B construct: RNRTL(pSer)VESL (crude synthetic peptide)

Average holdup BI: 0.17
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): 2.69
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: 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: ARHGEF2
Protein B fragment: 117-126
Protein B site: 14-3-3_pSer122
Protein B modification: (SEP)122
Protein B sequence: RERPSSAIYP
Protein B construct: RERPS(pSer)AIYP (crude synthetic peptide)

Average holdup BI: 0.33
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): 2.56
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
Measured pKd: 3.63


Experiment series 13

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: ARHGEF2
Protein B fragment: 881-890
Protein B site: 14-3-3_pSer886
Protein B modification: (SEP)886
Protein B sequence: DPRRRSLPAG
Protein B construct: DPRRR(pSer)LPAG (crude synthetic peptide)

Average holdup BI: 0.38
Holdup BI standard deviation: 0.13
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): 2.41
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: 3.74


Experiment series 14

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: ARHGEF2
Protein B fragment: 640-649
Protein B site: 14-3-3_pSer645
Protein B modification: (SEP)645
Protein B sequence: LFRSESLESP
Protein B construct: LFRSE(pSer)LESP (crude synthetic peptide)

Average holdup BI: 0.1
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): 2.18
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 construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: ARHGEF2
Protein B fragment: 706-715
Protein B site: 14-3-3_pSer711
Protein B modification: (SEP)711
Protein B sequence: RTFNGSIEL-
Protein B construct: ARTFNG(pSer)IEL (crude synthetic peptide)

Average holdup BI: -0.35
Holdup BI standard deviation: 0.29
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): 2.03
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 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: ARHGEF2
Protein B fragment: 640-649
Protein B site: 14-3-3_pSer645
Protein B modification: (SEP)645
Protein B sequence: LFRSESLESP
Protein B construct: LFRSE(pSer)LESP (crude synthetic peptide)

Average holdup BI: 0.13
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): 1.73
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
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: ARHGEF2
Protein B fragment: 117-126
Protein B site: 14-3-3_pSer122
Protein B modification: (SEP)122
Protein B sequence: RERPSSAIYP
Protein B construct: RERPS(pSer)AIYP (crude synthetic peptide)

Average holdup BI: 0.32
Holdup BI standard deviation: 0.2
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): 1.64
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: 3.62


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: ARHGEF2
Protein B fragment: 706-715
Protein B site: 14-3-3_pSer711
Protein B modification: (SEP)711
Protein B sequence: RTFNGSIEL-
Protein B construct: ARTFNG(pSer)IEL (crude synthetic peptide)

Average holdup BI: -0.28
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): 1.63
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 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: ARHGEF2
Protein B fragment: 172-181
Protein B site: 14-3-3_pSer177
Protein B modification: (SEP)177
Protein B sequence: SQSTDSLNMR
Protein B construct: SQSTD(pSer)LNMR (crude synthetic peptide)

Average holdup BI: 0.19
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): 1.46
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
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: ARHGEF2
Protein B fragment: 948-957
Protein B site: 14-3-3_pSer953
Protein B modification: (SEP)953
Protein B sequence: EEEGSSRLSP
Protein B construct: EEEGS(pSer)RLSP (crude synthetic peptide)

Average holdup BI: 0.14
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): 1.39
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: 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: ARHGEF2
Protein B fragment: 172-181
Protein B site: 14-3-3_pSer177
Protein B modification: (SEP)177
Protein B sequence: SQSTDSLNMR
Protein B construct: SQSTD(pSer)LNMR (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): 0.92
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 22

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: ARHGEF2
Protein B fragment: 181-190
Protein B site: 14-3-3_pSer186
Protein B modification: (SEP)186
Protein B sequence: RNRTLSVESL
Protein B construct: RNRTL(pSer)VESL (crude synthetic peptide)

Average holdup BI: 0.06
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): 0.89
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: 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: ARHGEF2
Protein B fragment: 167-176
Protein B site: 14-3-3_pSer172
Protein B modification: (SEP)172
Protein B sequence: LRRILSQSTD
Protein B construct: LRRIL(pSer)QSTD (crude synthetic peptide)

Average holdup BI: 0.04
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): 0.8
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 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: ARHGEF2
Protein B fragment: 936-945
Protein B site: 14-3-3_pSer941
Protein B modification: (SEP)941
Protein B sequence: RLQDSSDPDT
Protein B construct: RLQDS(pSer)DPDT (crude synthetic peptide)

Average holdup BI: -0.22
Holdup BI standard deviation: 0.42
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 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: ARHGEF2
Protein B fragment: 640-649
Protein B site: 14-3-3_pSer645
Protein B modification: (SEP)645
Protein B sequence: LFRSESLESP
Protein B construct: LFRSE(pSer)LESP (crude synthetic peptide)

Average holdup BI: 0.05
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): 0.73
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: 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: ARHGEF2
Protein B fragment: 128-137
Protein B site: 14-3-3_pSer133
Protein B modification: (SEP)133
Protein B sequence: DSFRQSLLGS
Protein B construct: DSFRQ(pSer)LLGS (crude synthetic peptide)

Average holdup BI: -0.03
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): 0.63
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: 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: ARHGEF2
Protein B fragment: 167-176
Protein B site: 14-3-3_pSer172
Protein B modification: (SEP)172
Protein B sequence: LRRILSQSTD
Protein B construct: LRRIL(pSer)QSTD (crude synthetic peptide)

Average holdup BI: 0.04
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): 0.63
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: ARHGEF2
Protein B fragment: 948-957
Protein B site: 14-3-3_pSer953
Protein B modification: (SEP)953
Protein B sequence: EEEGSSRLSP
Protein B construct: EEEGS(pSer)RLSP (crude synthetic peptide)

Average holdup BI: -0.03
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): 0.6
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 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: ARHGEF2
Protein B fragment: 936-945
Protein B site: 14-3-3_pSer941
Protein B modification: (SEP)941
Protein B sequence: RLQDSSDPDT
Protein B construct: RLQDS(pSer)DPDT (crude synthetic peptide)

Average holdup BI: -0.1
Holdup BI standard deviation: 0.18
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): 0.56
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 30

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: ARHGEF2
Protein B fragment: 714-723
Protein B site: 14-3-3_pSer719
Protein B modification: (SEP)719
Protein B sequence: LCRADSDSSQ
Protein B construct: LCRAD(pSer)DSSQ (crude synthetic peptide)

Average holdup BI: -0.17
Holdup BI standard deviation: 0.27
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): 0.4
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 31

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: ARHGEF2
Protein B fragment: 128-137
Protein B site: 14-3-3_pSer133
Protein B modification: (SEP)133
Protein B sequence: DSFRQSLLGS
Protein B construct: DSFRQ(pSer)LLGS (crude synthetic peptide)

Average holdup BI: -0.03
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): 0.31
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 32

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: ARHGEF2
Protein B fragment: 936-945
Protein B site: 14-3-3_pSer941
Protein B modification: (SEP)941
Protein B sequence: RLQDSSDPDT
Protein B construct: RLQDS(pSer)DPDT (crude synthetic peptide)

Average holdup BI: -0.03
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): 0.3
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: ARHGEF2
Protein B fragment: 167-176
Protein B site: 14-3-3_pSer172
Protein B modification: (SEP)172
Protein B sequence: LRRILSQSTD
Protein B construct: LRRIL(pSer)QSTD (crude synthetic peptide)

Average holdup BI: -0.04
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): 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: 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: ARHGEF2
Protein B fragment: 124-133
Protein B site: 14-3-3_pSer129
Protein B modification: (SEP)129
Protein B sequence: IYPSDSFRQS
Protein B construct: IYPSD(pSer)FRQS (crude synthetic peptide)

Average holdup BI: -0.01
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): 0.25
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B protein: ARHGEF2
Protein B fragment: 172-181
Protein B site: 14-3-3_pSer177
Protein B modification: (SEP)177
Protein B sequence: SQSTDSLNMR
Protein B construct: SQSTD(pSer)LNMR (crude synthetic peptide)

Average holdup BI: -0.09
Holdup BI standard deviation: 0.32
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 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: ARHGEF2
Protein B fragment: 124-133
Protein B site: 14-3-3_pSer129
Protein B modification: (SEP)129
Protein B sequence: IYPSDSFRQS
Protein B construct: IYPSD(pSer)FRQS (crude synthetic peptide)

Average holdup BI: -0.01
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): 0.19
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: ARHGEF2
Protein B fragment: 128-137
Protein B site: 14-3-3_pSer133
Protein B modification: (SEP)133
Protein B sequence: DSFRQSLLGS
Protein B construct: DSFRQ(pSer)LLGS (crude synthetic peptide)

Average holdup BI: 0.01
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): 0.13
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 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: ARHGEF2
Protein B fragment: 948-957
Protein B site: 14-3-3_pSer953
Protein B modification: (SEP)953
Protein B sequence: EEEGSSRLSP
Protein B construct: EEEGS(pSer)RLSP (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): 0.12
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 39

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: ARHGEF2
Protein B fragment: 124-133
Protein B site: 14-3-3_pSer129
Protein B modification: (SEP)129
Protein B sequence: IYPSDSFRQS
Protein B construct: IYPSD(pSer)FRQS (crude synthetic peptide)

Average holdup BI: 0
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): 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 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: ARHGEF2
Protein B fragment: 706-715
Protein B site: 14-3-3_pSer711
Protein B modification: (SEP)711
Protein B sequence: RTFNGSIEL-
Protein B construct: ARTFNG(pSer)IEL (crude synthetic peptide)

Average holdup BI: 0
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): 0.02
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 41

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: ARHGEF2
Protein B fragment: 138-147
Protein B site: 14-3-3_pSer143
Protein B modification: (SEP)143
Protein B sequence: RRGRSSLSLA
Protein B construct: RRGRS(pSer)LSLA (crude synthetic peptide)

Average holdup BI: 0
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): 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.