Details of experiments between P61981 and Q9Y2U5
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: MAP3K2
Protein B fragment: 234-243
Protein B site: 14-3-3_pSer239
Protein B modification: (SEP)239
Protein B sequence: MPRAQSYPDN
Protein B construct: MPRAQ(pSer)YPDN (crude synthetic peptide)

Average holdup BI: 0.85
Holdup BI standard deviation: 0.01
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): 11.68
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: 4.63


Experiment series 2

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: MAP3K2
Protein B fragment: 278-287
Protein B site: 14-3-3_pThr283
Protein B modification: (TPO)283
Protein B sequence: NDGRKTFPRA
Protein B construct: NDGRK(pThr)FPRA (crude synthetic peptide)

Average holdup BI: 0.78
Holdup BI standard deviation: 0.02
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): 8.7
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: 4.44


Experiment series 3

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: MAP3K2
Protein B fragment: 258-267
Protein B site: 14-3-3_pThr263
Protein B modification: (TPO)263
Protein B sequence: FGKGGTYPRR
Protein B construct: FGKGG(pThr)YPRR (crude synthetic peptide)

Average holdup BI: 0.89
Holdup BI standard deviation: 0.02
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): 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: 4.86


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: MAP3K2
Protein B fragment: 258-267
Protein B site: 14-3-3_pThr263
Protein B modification: (TPO)263
Protein B sequence: FGKGGTYPRR
Protein B construct: FGKGG(pThr)YPRR (crude synthetic peptide)

Average holdup BI: 0.93
Holdup BI standard deviation: 0.01
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): 7.62
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: 5.04


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: MAP3K2
Protein B fragment: 517-526
Protein B site: 14-3-3_pThr522
Protein B modification: (TPO)522
Protein B sequence: GMKSVTGTPY
Protein B construct: GMKSV(pThr)GTPY (crude synthetic peptide)

Average holdup BI: 0.73
Holdup BI standard deviation: 0.02
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): 7.2
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
Measured pKd: 4.32


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: MAP3K2
Protein B fragment: 234-243
Protein B site: 14-3-3_pSer239
Protein B modification: (SEP)239
Protein B sequence: MPRAQSYPDN
Protein B construct: MPRAQ(pSer)YPDN (crude synthetic peptide)

Average holdup BI: 0.87
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): 6.28
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
Measured pKd: 4.69


Experiment series 7

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: MAP3K2
Protein B fragment: 234-243
Protein B site: 14-3-3_pSer239
Protein B modification: (SEP)239
Protein B sequence: MPRAQSYPDN
Protein B construct: MPRAQ(pSer)YPDN (crude synthetic peptide)

Average holdup BI: 0.81
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): 5.94
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: 4.6


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: MAP3K2
Protein B fragment: 278-287
Protein B site: 14-3-3_pThr283
Protein B modification: (TPO)283
Protein B sequence: NDGRKTFPRA
Protein B construct: NDGRK(pThr)FPRA (crude synthetic peptide)

Average holdup BI: 0.72
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): 5.91
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: 4.28


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: MAP3K2
Protein B fragment: 258-267
Protein B site: 14-3-3_pThr263
Protein B modification: (TPO)263
Protein B sequence: FGKGGTYPRR
Protein B construct: FGKGG(pThr)YPRR (crude synthetic peptide)

Average holdup BI: 0.92
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): 5.65
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: 4.91


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: MAP3K2
Protein B fragment: 517-526
Protein B site: 14-3-3_pThr522
Protein B modification: (TPO)522
Protein B sequence: GMKSVTGTPY
Protein B construct: GMKSV(pThr)GTPY (crude synthetic peptide)

Average holdup BI: 0.56
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): 4.59
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: 3.98


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: MAP3K2
Protein B fragment: 517-526
Protein B site: 14-3-3_pThr522
Protein B modification: (TPO)522
Protein B sequence: GMKSVTGTPY
Protein B construct: GMKSV(pThr)GTPY (crude synthetic peptide)

Average holdup BI: 0.42
Holdup BI standard deviation: 0.06
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): 4.31
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
Measured pKd: 3.81


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: MAP3K2
Protein B fragment: 278-287
Protein B site: 14-3-3_pThr283
Protein B modification: (TPO)283
Protein B sequence: NDGRKTFPRA
Protein B construct: NDGRK(pThr)FPRA (crude synthetic peptide)

Average holdup BI: 0.64
Holdup BI standard deviation: 0.08
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): 3.15
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: 4.19


Experiment series 13

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: MAP3K2
Protein B fragment: 488-497
Protein B site: 14-3-3_pSer493
Protein B modification: (SEP)493
Protein B sequence: NILRDSTGNV
Protein B construct: NILRD(pSer)TGNV (crude synthetic peptide)

Average holdup BI: 0.03
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): 1.4
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: 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: MAP3K2
Protein B fragment: 243-252
Protein B site: 14-3-3_pSer248
Protein B modification: (SEP)248
Protein B sequence: NHQEFSDYDN
Protein B construct: NHQEF(pSer)DYDN (crude synthetic peptide)

Average holdup BI: -0.26
Holdup BI standard deviation: 0.38
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): 1.02
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 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: MAP3K2
Protein B fragment: 148-157
Protein B site: 14-3-3_pSer153
Protein B modification: (SEP)153
Protein B sequence: RKKRLSIIGP
Protein B construct: RKKRL(pSer)IIGP (crude synthetic peptide)

Average holdup BI: -0.25
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): 0.95
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
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 construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: MAP3K2
Protein B fragment: 148-157
Protein B site: 14-3-3_pSer153
Protein B modification: (SEP)153
Protein B sequence: RKKRLSIIGP
Protein B construct: RKKRL(pSer)IIGP (crude synthetic peptide)

Average holdup BI: 0.11
Holdup BI standard deviation: 0.13
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): 0.91
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: 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: MAP3K2
Protein B fragment: 515-524
Protein B site: 14-3-3_pSer520
Protein B modification: (SEP)520
Protein B sequence: GTGMKSVTGT
Protein B construct: GTGMK(pSer)VTGT (crude synthetic peptide)

Average holdup BI: 0.14
Holdup BI standard deviation: 0.16
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): 0.86
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
The affinity was below the detection threshold of the assay.


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: MAP3K2
Protein B fragment: 488-497
Protein B site: 14-3-3_pSer493
Protein B modification: (SEP)493
Protein B sequence: NILRDSTGNV
Protein B construct: NILRD(pSer)TGNV (crude synthetic peptide)

Average holdup BI: 0.01
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): 0.36
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 19

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: MAP3K2
Protein B fragment: 515-524
Protein B site: 14-3-3_pSer520
Protein B modification: (SEP)520
Protein B sequence: GTGMKSVTGT
Protein B construct: GTGMK(pSer)VTGT (crude synthetic peptide)

Average holdup BI: 0.02
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): 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: 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: MAP3K2
Protein B fragment: 243-252
Protein B site: 14-3-3_pSer248
Protein B modification: (SEP)248
Protein B sequence: NHQEFSDYDN
Protein B construct: NHQEF(pSer)DYDN (crude synthetic peptide)

Average holdup BI: 0.07
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): 0.14
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 21

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: MAP3K2
Protein B fragment: 515-524
Protein B site: 14-3-3_pSer520
Protein B modification: (SEP)520
Protein B sequence: GTGMKSVTGT
Protein B construct: GTGMK(pSer)VTGT (crude synthetic peptide)

Average holdup BI: 0.08
Holdup BI standard deviation: 0.48
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 22

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: MAP3K2
Protein B fragment: 488-497
Protein B site: 14-3-3_pSer493
Protein B modification: (SEP)493
Protein B sequence: NILRDSTGNV
Protein B construct: NILRD(pSer)TGNV (crude synthetic peptide)

Average holdup BI: 0
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): 0.03
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.