Details of experiments between P62258 and O75122
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 355-364
Protein B site: 14-3-3_pSer360
Protein B modification: (SEP)360
Protein B sequence: SSKASSLPGS
Protein B construct: SSKAS(pSer)LPGS (crude synthetic peptide)

Average holdup BI: 0.56
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.57
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: 3.95


Experiment series 2

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 450-459
Protein B site: 14-3-3_pSer455
Protein B modification: (SEP)455
Protein B sequence: KMVSQSQPGS
Protein B construct: KMVSQ(pSer)QPGS (crude synthetic peptide)

Average holdup BI: 0.44
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.55
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
Measured pKd: 3.74


Experiment series 3

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 355-364
Protein B site: 14-3-3_pSer360
Protein B modification: (SEP)360
Protein B sequence: SSKASSLPGS
Protein B construct: SSKAS(pSer)LPGS (crude synthetic peptide)

Average holdup BI: 0.49
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.08
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
Measured pKd: 3.92


Experiment series 4

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 450-459
Protein B site: 14-3-3_pSer455
Protein B modification: (SEP)455
Protein B sequence: KMVSQSQPGS
Protein B construct: KMVSQ(pSer)QPGS (crude synthetic peptide)

Average holdup BI: 0.5
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.36
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: 3.94


Experiment series 5

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 425-434
Protein B site: 14-3-3_pSer430
Protein B modification: (SEP)430
Protein B sequence: VRAKLSAPLA
Protein B construct: VRAKL(pSer)APLA (crude synthetic peptide)

Average holdup BI: 0.12
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.19
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 6

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 1048-1057
Protein B site: 14-3-3_pSer1053
Protein B modification: (SEP)1053
Protein B sequence: FPDDLSLDHS
Protein B construct: FPDDL(pSer)LDHS (crude synthetic peptide)

Average holdup BI: -0.12
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.87
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 347-356
Protein B site: 14-3-3_pSer352
Protein B modification: (SEP)352
Protein B sequence: VAGRVSAGSS
Protein B construct: VAGRV(pSer)AGSS (crude synthetic peptide)

Average holdup BI: -0.34
Holdup BI standard deviation: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 143
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: 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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 425-434
Protein B site: 14-3-3_pSer430
Protein B modification: (SEP)430
Protein B sequence: VRAKLSAPLA
Protein B construct: VRAKL(pSer)APLA (crude synthetic peptide)

Average holdup BI: 0.12
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.59
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 9

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 359-368
Protein B site: 14-3-3_pSer364
Protein B modification: (SEP)364
Protein B sequence: SSLPGSLQRS
Protein B construct: SSLPG(pSer)LQRS (crude synthetic peptide)

Average holdup BI: 0.05
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.15
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 947-956
Protein B site: 14-3-3_pSer952
Protein B modification: (SEP)952
Protein B sequence: AIQNFSFRSQ
Protein B construct: AIQNF(pSer)FRSQ (crude synthetic peptide)

Average holdup BI: -0.09
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.04
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 11

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 365-374
Protein B site: 14-3-3_pSer370
Protein B modification: (SEP)370
Protein B sequence: LQRSRSDIDV
Protein B construct: LQRSR(pSer)DIDV (crude synthetic peptide)

Average holdup BI: 0.12
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 143
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: 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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 3-12
Protein B site: 14-3-3_pSer8
Protein B modification: (SEP)8
Protein B sequence: MGDDKSFDDE
Protein B construct: MGDDK(pSer)FDDE (crude synthetic peptide)

Average holdup BI: -0.46
Holdup BI standard deviation: 0.98
Immobilized partner concentration in holdup experiment (10-6M): 143
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 13

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 591-600
Protein B site: 14-3-3_pSer596
Protein B modification: (SEP)596
Protein B sequence: VLNTGSDVEE
Protein B construct: VLNTG(pSer)DVEE (crude synthetic peptide)

Average holdup BI: -0.15
Holdup BI standard deviation: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 143
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 14

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 1029-1038
Protein B site: 14-3-3_pSer1034
Protein B modification: (SEP)1034
Protein B sequence: SPFNKSALKE
Protein B construct: SPFNK(pSer)ALKE (crude synthetic peptide)

Average holdup BI: 0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.69
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 15

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 639-648
Protein B site: 14-3-3_pSer644
Protein B modification: (SEP)644
Protein B sequence: SSRNGSIPTY
Protein B construct: SSRNG(pSer)IPTY (crude synthetic peptide)

Average holdup BI: 0.05
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.65
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 16

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 1029-1038
Protein B site: 14-3-3_pSer1034
Protein B modification: (SEP)1034
Protein B sequence: SPFNKSALKE
Protein B construct: SPFNK(pSer)ALKE (crude synthetic peptide)

Average holdup BI: -0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.5
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 365-374
Protein B site: 14-3-3_pSer370
Protein B modification: (SEP)370
Protein B sequence: LQRSRSDIDV
Protein B construct: LQRSR(pSer)DIDV (crude synthetic peptide)

Average holdup BI: 0.24
Holdup BI standard deviation: 0.35
Immobilized partner concentration in holdup experiment (10-6M): 116
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: 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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 1007-1016
Protein B site: 14-3-3_pSer1012
Protein B modification: (SEP)1012
Protein B sequence: SMPTHSSPRS
Protein B construct: SMPTH(pSer)SPRS (crude synthetic peptide)

Average holdup BI: 0.22
Holdup BI standard deviation: 0.4
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.45
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 484-493
Protein B site: 14-3-3_pSer489
Protein B modification: (SEP)489
Protein B sequence: LVNSASAQKR
Protein B construct: LVNSA(pSer)AQKR (crude synthetic peptide)

Average holdup BI: -0.19
Holdup BI standard deviation: 0.49
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.33
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 947-956
Protein B site: 14-3-3_pSer952
Protein B modification: (SEP)952
Protein B sequence: AIQNFSFRSQ
Protein B construct: AIQNF(pSer)FRSQ (crude synthetic peptide)

Average holdup BI: 0.03
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.24
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 1048-1057
Protein B site: 14-3-3_pSer1053
Protein B modification: (SEP)1053
Protein B sequence: FPDDLSLDHS
Protein B construct: FPDDL(pSer)LDHS (crude synthetic peptide)

Average holdup BI: 0.02
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.18
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 22

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 1007-1016
Protein B site: 14-3-3_pSer1012
Protein B modification: (SEP)1012
Protein B sequence: SMPTHSSPRS
Protein B construct: SMPTH(pSer)SPRS (crude synthetic peptide)

Average holdup BI: -0.05
Holdup BI standard deviation: 0.24
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.15
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 23

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 347-356
Protein B site: 14-3-3_pSer352
Protein B modification: (SEP)352
Protein B sequence: VAGRVSAGSS
Protein B construct: VAGRV(pSer)AGSS (crude synthetic peptide)

Average holdup BI: -0.02
Holdup BI standard deviation: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 116
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 24

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 639-648
Protein B site: 14-3-3_pSer644
Protein B modification: (SEP)644
Protein B sequence: SSRNGSIPTY
Protein B construct: SSRNG(pSer)IPTY (crude synthetic peptide)

Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 116
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 25

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 591-600
Protein B site: 14-3-3_pSer596
Protein B modification: (SEP)596
Protein B sequence: VLNTGSDVEE
Protein B construct: VLNTG(pSer)DVEE (crude synthetic peptide)

Average holdup BI: 0.02
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 116
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 611-620
Protein B site: 14-3-3_pSer616
Protein B modification: (SEP)616
Protein B sequence: RRRYESYGMH
Protein B construct: RRRYE(pSer)YGMH (crude synthetic peptide)

Average holdup BI: -0.02
Holdup BI standard deviation: 0.29
Immobilized partner concentration in holdup experiment (10-6M): 116
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 27

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 950-959
Protein B site: 14-3-3_pSer955
Protein B modification: (SEP)955
Protein B sequence: NFSFRSQEDM
Protein B construct: NFSFR(pSer)QEDM (crude synthetic peptide)

Average holdup BI: 0.01
Holdup BI standard deviation: 0.18
Immobilized partner concentration in holdup experiment (10-6M): 143
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 28

Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B protein: CLASP2
Protein B fragment: 359-368
Protein B site: 14-3-3_pSer364
Protein B modification: (SEP)364
Protein B sequence: SSLPGSLQRS
Protein B construct: SSLPG(pSer)LQRS (crude synthetic peptide)

Average holdup BI: 0
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 116
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.