Details of experiments between P61981 and Q7Z460
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 1079-1088
Protein B site: 14-3-3_pThr1084
Protein B modification: (TPO)1084
Protein B sequence: RTSPLTSPTN
Protein B construct: RTSPL(pThr)SPTN (crude synthetic peptide)
Average holdup BI: 0.77
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 12.53
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: 4.41
Experiment series 2
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 1079-1088
Protein B site: 14-3-3_pThr1084
Protein B modification: (TPO)1084
Protein B sequence: RTSPLTSPTN
Protein B construct: RTSPL(pThr)SPTN (crude synthetic peptide)
Average holdup BI: 0.7
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 12.42
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
Measured pKd: 4.33
Experiment series 3
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 1079-1088
Protein B site: 14-3-3_pThr1084
Protein B modification: (TPO)1084
Protein B sequence: RTSPLTSPTN
Protein B construct: RTSPL(pThr)SPTN (crude synthetic peptide)
Average holdup BI: 0.75
Holdup BI standard deviation: 0.01
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): 10.95
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: 4.35
Experiment series 4
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 682-691
Protein B site: 14-3-3_pSer687
Protein B modification: (SEP)687
Protein B sequence: KVVSQSQPGS
Protein B construct: KVVSQ(pSer)QPGS (crude synthetic peptide)
Average holdup BI: 0.82
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.62
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: 4.53
Experiment series 5
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 277-286
Protein B site: 14-3-3_pThr282
Protein B modification: (TPO)282
Protein B sequence: RLGSSTLGSK
Protein B construct: RLGSS(pThr)LGSK (crude synthetic peptide)
Average holdup BI: 0.66
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): 6.51
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: 4.16
Experiment series 6
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 277-286
Protein B site: 14-3-3_pThr282
Protein B modification: (TPO)282
Protein B sequence: RLGSSTLGSK
Protein B construct: RLGSS(pThr)LGSK (crude synthetic peptide)
Average holdup BI: 0.74
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.38
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: 4.33
Experiment series 7
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 631-640
Protein B site: 14-3-3_pSer636
Protein B modification: (SEP)636
Protein B sequence: PGSYASLGRI
Protein B construct: PGSYA(pSer)LGRI (crude synthetic peptide)
Average holdup BI: 0.51
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.96
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: 3.91
Experiment series 8
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 631-640
Protein B site: 14-3-3_pSer636
Protein B modification: (SEP)636
Protein B sequence: PGSYASLGRI
Protein B construct: PGSYA(pSer)LGRI (crude synthetic peptide)
Average holdup BI: 0.49
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): 5.95
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: 3.85
Experiment series 9
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 682-691
Protein B site: 14-3-3_pSer687
Protein B modification: (SEP)687
Protein B sequence: KVVSQSQPGS
Protein B construct: KVVSQ(pSer)QPGS (crude synthetic peptide)
Average holdup BI: 0.76
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): 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: 4.38
Experiment series 10
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 589-598
Protein B site: 14-3-3_pSer594
Protein B modification: (SEP)594
Protein B sequence: VSTTGSLQRS
Protein B construct: VSTTG(pSer)LQRS (crude synthetic peptide)
Average holdup BI: 0.22
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.16
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 11
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 631-640
Protein B site: 14-3-3_pSer636
Protein B modification: (SEP)636
Protein B sequence: PGSYASLGRI
Protein B construct: PGSYA(pSer)LGRI (crude synthetic peptide)
Average holdup BI: 0.53
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.51
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
Experiment series 12
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 682-691
Protein B site: 14-3-3_pSer687
Protein B modification: (SEP)687
Protein B sequence: KVVSQSQPGS
Protein B construct: KVVSQ(pSer)QPGS (crude synthetic peptide)
Average holdup BI: 0.69
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.5
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
Measured pKd: 4.3
Experiment series 13
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 589-598
Protein B site: 14-3-3_pSer594
Protein B modification: (SEP)594
Protein B sequence: VSTTGSLQRS
Protein B construct: VSTTG(pSer)LQRS (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.32
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 589-598
Protein B site: 14-3-3_pSer594
Protein B modification: (SEP)594
Protein B sequence: VSTTGSLQRS
Protein B construct: VSTTG(pSer)LQRS (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.09
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): 2.1
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 15
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 792-801
Protein B site: 14-3-3_pSer797
Protein B modification: (SEP)797
Protein B sequence: RVLSTSTDLE
Protein B construct: RVLST(pSer)TDLE (crude synthetic peptide)
Average holdup BI: 0.04
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): 1.96
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
The affinity was below the detection threshold of the assay.
Experiment series 16
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 1191-1200
Protein B site: 14-3-3_pSer1196
Protein B modification: (SEP)1196
Protein B sequence: KFSFRSQEDL
Protein B construct: KFSFR(pSer)QEDL (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 115
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: 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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 277-286
Protein B site: 14-3-3_pThr282
Protein B modification: (TPO)282
Protein B sequence: RLGSSTLGSK
Protein B construct: RLGSS(pThr)LGSK (crude synthetic peptide)
Average holdup BI: 0.61
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.86
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: 4.15
Experiment series 18
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 1191-1200
Protein B site: 14-3-3_pSer1196
Protein B modification: (SEP)1196
Protein B sequence: KFSFRSQEDL
Protein B construct: KFSFR(pSer)QEDL (crude synthetic peptide)
Average holdup BI: -0.29
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): 1.77
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 19
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 792-801
Protein B site: 14-3-3_pSer797
Protein B modification: (SEP)797
Protein B sequence: RVLSTSTDLE
Protein B construct: RVLST(pSer)TDLE (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.75
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 593-602
Protein B site: 14-3-3_pSer598
Protein B modification: (SEP)598
Protein B sequence: GSLQRSRSDI
Protein B construct: GSLQR(pSer)RSDI (crude synthetic peptide)
Average holdup BI: 0.23
Holdup BI standard deviation: 0.17
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.32
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 790-799
Protein B site: 14-3-3_pSer795
Protein B modification: (SEP)795
Protein B sequence: AMRVLSTSTD
Protein B construct: AMRVL(pSer)TSTD (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.14
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): 1.31
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 22
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 628-637
Protein B site: 14-3-3_pSer633
Protein B modification: (SEP)633
Protein B sequence: ALPPGSYASL
Protein B construct: ALPPG(pSer)YASL (crude synthetic peptide)
Average holdup BI: 0.34
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): 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 23
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 1191-1200
Protein B site: 14-3-3_pSer1196
Protein B modification: (SEP)1196
Protein B sequence: KFSFRSQEDL
Protein B construct: KFSFR(pSer)QEDL (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.18
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.83
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 792-801
Protein B site: 14-3-3_pSer797
Protein B modification: (SEP)797
Protein B sequence: RVLSTSTDLE
Protein B construct: RVLST(pSer)TDLE (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.61
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: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 593-602
Protein B site: 14-3-3_pSer598
Protein B modification: (SEP)598
Protein B sequence: GSLQRSRSDI
Protein B construct: GSLQR(pSer)RSDI (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.07
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): 0.58
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
The affinity was below the detection threshold of the assay.
Experiment series 26
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 563-572
Protein B site: 14-3-3_pSer568
Protein B modification: (SEP)568
Protein B sequence: LNRPLSAKRS
Protein B construct: LNRPL(pSer)AKRS (crude synthetic peptide)
Average holdup BI: -0.1
Holdup BI standard deviation: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 115
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 27
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 790-799
Protein B site: 14-3-3_pSer795
Protein B modification: (SEP)795
Protein B sequence: AMRVLSTSTD
Protein B construct: AMRVL(pSer)TSTD (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.33
Immobilized partner concentration in holdup experiment (10-6M): 115
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 28
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 718-727
Protein B site: 14-3-3_pSer723
Protein B modification: (SEP)723
Protein B sequence: SKIPRSQGCS
Protein B construct: SKIPR(pSer)QGCS (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.38
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.28
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 29
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 563-572
Protein B site: 14-3-3_pSer568
Protein B modification: (SEP)568
Protein B sequence: LNRPLSAKRS
Protein B construct: LNRPL(pSer)AKRS (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.59
Immobilized partner concentration in holdup experiment (10-6M): 129
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 30
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 790-799
Protein B site: 14-3-3_pSer795
Protein B modification: (SEP)795
Protein B sequence: AMRVLSTSTD
Protein B construct: AMRVL(pSer)TSTD (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 129
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 31
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 628-637
Protein B site: 14-3-3_pSer633
Protein B modification: (SEP)633
Protein B sequence: ALPPGSYASL
Protein B construct: ALPPG(pSer)YASL (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 1.02
Immobilized partner concentration in holdup experiment (10-6M): 115
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 32
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 563-572
Protein B site: 14-3-3_pSer568
Protein B modification: (SEP)568
Protein B sequence: LNRPLSAKRS
Protein B construct: LNRPL(pSer)AKRS (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.65
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): 0.06
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 33
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 628-637
Protein B site: 14-3-3_pSer633
Protein B modification: (SEP)633
Protein B sequence: ALPPGSYASL
Protein B construct: ALPPG(pSer)YASL (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 129
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 34
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 593-602
Protein B site: 14-3-3_pSer598
Protein B modification: (SEP)598
Protein B sequence: GSLQRSRSDI
Protein B construct: GSLQR(pSer)RSDI (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 129
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.
Experiment series 35
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 682-691
Protein B site: 14-3-3_pSer687
Protein B modification: (SEP)687
Protein B sequence: KVVSQSQPGS
Protein B construct: biotin-ttds-KVVSQ(pSer)QPGS (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 48
Standard deviation of dissociation constant (10-6M): 4.5
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022.01.18
Number of measurements: 3
Measured pKd: 4.32
Experiment series 36
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: CLASP1
Protein B fragment: 682-691
Protein B site: 14-3-3_pSer687
Protein B modification: (SEP)687
Protein B sequence: KVVSQSQPGS
Protein B construct: biotin-ttds-KVVSQ(pSer)QPGS (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 25
Standard deviation of dissociation constant (10-6M): 4
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022.01.10
Number of measurements: 3
Measured pKd: 4.42