Details of experiments between Q8IZ21 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: PHACTR4
Protein A fragment: 459-468
Protein A site: 14-3-3_pSer464
Protein A modification: (SEP)464
Protein A sequence: LGRTRSLPIT
Protein A construct: LGRTR(pSer)LPIT (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: 0.9
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): 7.78
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: 4.93


Experiment series 2

Protein A protein: PHACTR4
Protein A fragment: 459-468
Protein A site: 14-3-3_pSer464
Protein A modification: (SEP)464
Protein A sequence: LGRTRSLPIT
Protein A construct: LGRTR(pSer)LPIT (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: 0.92
Holdup BI standard deviation: 0.01
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): 6.96
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


Experiment series 3

Protein A protein: PHACTR4
Protein A fragment: 459-468
Protein A site: 14-3-3_pSer464
Protein A modification: (SEP)464
Protein A sequence: LGRTRSLPIT
Protein A construct: LGRTR(pSer)LPIT (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: 0.94
Holdup BI standard deviation: 0.01
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): 6.88
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: 5.04


Experiment series 4

Protein A protein: PHACTR4
Protein A fragment: 265-274
Protein A site: 14-3-3_pSer270
Protein A modification: (SEP)270
Protein A sequence: PAHRNSNPVI
Protein A construct: PAHRN(pSer)NPVI (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: 0.1
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): 1.42
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: PHACTR4
Protein A fragment: 126-135
Protein A site: 14-3-3_pSer131
Protein A modification: (SEP)131
Protein A sequence: PVRLASLRKA
Protein A construct: PVRLA(pSer)LRKA (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: 0.13
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): 1.17
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 6

Protein A protein: PHACTR4
Protein A fragment: 65-74
Protein A site: 14-3-3_pSer70
Protein A modification: (SEP)70
Protein A sequence: LERKISMRKP
Protein A construct: LERKI(pSer)MRKP (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: 0.09
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): 1.17
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 7

Protein A protein: PHACTR4
Protein A fragment: 126-135
Protein A site: 14-3-3_pSer131
Protein A modification: (SEP)131
Protein A sequence: PVRLASLRKA
Protein A construct: PVRLA(pSer)LRKA (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): 0.98
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 8

Protein A protein: PHACTR4
Protein A fragment: 142-151
Protein A site: 14-3-3_pSer147
Protein A modification: (SEP)147
Protein A sequence: KKRLGSTGSQ
Protein A construct: KKRLG(pSer)TGSQ (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: -0.45
Holdup BI standard deviation: 0.41
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): 0.92
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
The affinity was below the detection threshold of the assay.


Experiment series 9

Protein A protein: PHACTR4
Protein A fragment: 623-632
Protein A site: 14-3-3_pSer628
Protein A modification: (SEP)628
Protein A sequence: LTRKLSQRPT
Protein A construct: LTRKL(pSer)QRPT (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: 0.25
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): 0.77
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: PHACTR4
Protein A fragment: 585-594
Protein A site: 14-3-3_pSer590
Protein A modification: (SEP)590
Protein A sequence: LIRRLSQRPT
Protein A construct: LIRRL(pSer)QRPT (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: -0.08
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): 0.77
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 11

Protein A protein: PHACTR4
Protein A fragment: 585-594
Protein A site: 14-3-3_pSer590
Protein A modification: (SEP)590
Protein A sequence: LIRRLSQRPT
Protein A construct: LIRRL(pSer)QRPT (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: 0.1
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): 0.74
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 12

Protein A protein: PHACTR4
Protein A fragment: 142-151
Protein A site: 14-3-3_pSer147
Protein A modification: (SEP)147
Protein A sequence: KKRLGSTGSQ
Protein A construct: KKRLG(pSer)TGSQ (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: 0.33
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): 0.72
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: PHACTR4
Protein A fragment: 265-274
Protein A site: 14-3-3_pSer270
Protein A modification: (SEP)270
Protein A sequence: PAHRNSNPVI
Protein A construct: PAHRN(pSer)NPVI (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: 0.12
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 14

Protein A protein: PHACTR4
Protein A fragment: 112-121
Protein A site: 14-3-3_pSer117
Protein A modification: (SEP)117
Protein A sequence: GNARSSSPVQ
Protein A construct: GNARS(pSer)SPVQ (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: 0.26
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): 0.43
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: PHACTR4
Protein A fragment: 585-594
Protein A site: 14-3-3_pSer590
Protein A modification: (SEP)590
Protein A sequence: LIRRLSQRPT
Protein A construct: LIRRL(pSer)QRPT (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: -0.06
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): 0.37
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: PHACTR4
Protein A fragment: 265-274
Protein A site: 14-3-3_pSer270
Protein A modification: (SEP)270
Protein A sequence: PAHRNSNPVI
Protein A construct: PAHRN(pSer)NPVI (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: -0.08
Holdup BI standard deviation: 0.19
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): 0.36
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 17

Protein A protein: PHACTR4
Protein A fragment: 552-561
Protein A site: 14-3-3_pSer557
Protein A modification: (SEP)557
Protein A sequence: LNHRPSEPEL
Protein A construct: LNHRP(pSer)EPEL (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: 0.06
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): 0.23
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 18

Protein A protein: PHACTR4
Protein A fragment: 142-151
Protein A site: 14-3-3_pSer147
Protein A modification: (SEP)147
Protein A sequence: KKRLGSTGSQ
Protein A construct: KKRLG(pSer)TGSQ (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: 0.16
Holdup BI standard deviation: 0.36
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 19

Protein A protein: PHACTR4
Protein A fragment: 112-121
Protein A site: 14-3-3_pSer117
Protein A modification: (SEP)117
Protein A sequence: GNARSSSPVQ
Protein A construct: GNARS(pSer)SPVQ (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: 0.76
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): 0.2
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 20

Protein A protein: PHACTR4
Protein A fragment: 623-632
Protein A site: 14-3-3_pSer628
Protein A modification: (SEP)628
Protein A sequence: LTRKLSQRPT
Protein A construct: LTRKL(pSer)QRPT (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: 0.33
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): 0.16
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 21

Protein A protein: PHACTR4
Protein A fragment: 65-74
Protein A site: 14-3-3_pSer70
Protein A modification: (SEP)70
Protein A sequence: LERKISMRKP
Protein A construct: LERKI(pSer)MRKP (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: 0.08
Holdup BI standard deviation: 0.36
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): 0.16
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: PHACTR4
Protein A fragment: 552-561
Protein A site: 14-3-3_pSer557
Protein A modification: (SEP)557
Protein A sequence: LNHRPSEPEL
Protein A construct: LNHRP(pSer)EPEL (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: 0.09
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 23

Protein A protein: PHACTR4
Protein A fragment: 126-135
Protein A site: 14-3-3_pSer131
Protein A modification: (SEP)131
Protein A sequence: PVRLASLRKA
Protein A construct: PVRLA(pSer)LRKA (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: 0.03
Holdup BI standard deviation: 0.19
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): 0.13
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: PHACTR4
Protein A fragment: 112-121
Protein A site: 14-3-3_pSer117
Protein A modification: (SEP)117
Protein A sequence: GNARSSSPVQ
Protein A construct: GNARS(pSer)SPVQ (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: 0.44
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 25

Protein A protein: PHACTR4
Protein A fragment: 65-74
Protein A site: 14-3-3_pSer70
Protein A modification: (SEP)70
Protein A sequence: LERKISMRKP
Protein A construct: LERKI(pSer)MRKP (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: -0.05
Holdup BI standard deviation: 0.47
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 26

Protein A protein: PHACTR4
Protein A fragment: 552-561
Protein A site: 14-3-3_pSer557
Protein A modification: (SEP)557
Protein A sequence: LNHRPSEPEL
Protein A construct: LNHRP(pSer)EPEL (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: 0.07
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): 0.06
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 27

Protein A protein: PHACTR4
Protein A fragment: 623-632
Protein A site: 14-3-3_pSer628
Protein A modification: (SEP)628
Protein A sequence: LTRKLSQRPT
Protein A construct: LTRKL(pSer)QRPT (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: 0.31
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 28

Protein A protein: PHACTR4
Protein A fragment: 438-447
Protein A site: 14-3-3_pSer443
Protein A modification: (SEP)443
Protein A sequence: SHRAHSLLFE
Protein A construct: SHRAH(pSer)LLFE (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: 0.15
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): 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 29

Protein A protein: PHACTR4
Protein A fragment: 438-447
Protein A site: 14-3-3_pSer443
Protein A modification: (SEP)443
Protein A sequence: SHRAHSLLFE
Protein A construct: SHRAH(pSer)LLFE (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: -0.04
Holdup BI standard deviation: 1.06
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): 0.02
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.