Details of experiments between P31946 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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.7
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 9.18
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: 4.36
Experiment series 2
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.67
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.76
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: 4.29
Experiment series 3
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.7
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.96
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: 4.24
Experiment series 4
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.71
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.24
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: 4.26
Experiment series 5
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.37
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.64
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: 3.64
Experiment series 6
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.33
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.07
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: 3.69
Experiment series 7
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.29
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.85
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: 3.6
Experiment series 8
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.5
Holdup BI standard deviation: 0.17
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.84
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: 3.88
Experiment series 9
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.08
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.1
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.47
Holdup BI standard deviation: 0.21
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.06
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
Measured pKd: 3.82
Experiment series 11
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.12
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.56
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 12
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.36
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
The affinity was below the detection threshold of the assay.
Experiment series 13
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.35
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 14
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.07
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.21
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.11
Holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 102
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 16
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.13
Holdup BI standard deviation: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.52
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.14
Holdup BI standard deviation: 0.3
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.42
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 18
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: -0.11
Holdup BI standard deviation: 0.24
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.4
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 19
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: -0.06
Holdup BI standard deviation: 0.17
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.29
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.01
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.22
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 21
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: -0.05
Holdup BI standard deviation: 0.19
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.2
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.04
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 102
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: 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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.01
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 102
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: 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 24
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.06
Holdup BI standard deviation: 0.28
Immobilized partner concentration in holdup experiment (10-6M): 102
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: 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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: -0.06
Holdup BI standard deviation: 0.33
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.17
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 26
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: -0.11
Holdup BI standard deviation: 0.77
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.11
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: 0.01
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 28
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: -0.12
Holdup BI standard deviation: 0.9
Immobilized partner concentration in holdup experiment (10-6M): 102
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: 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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
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: -0.03
Holdup BI standard deviation: 0.43
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.05
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.