Details of experiments between P61981 and Q99759
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: MAP3K3
Protein B fragment: 245-254
Protein B site: 14-3-3_pSer250
Protein B modification: (SEP)250
Protein B sequence: MSRAQSFPDN
Protein B construct: MSRAQ(pSer)FPDN (crude synthetic peptide)

Average holdup BI: 0.81
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): 13.69
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
Measured pKd: 4.51


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: MAP3K3
Protein B fragment: 352-361
Protein B site: 14-3-3_pSer357
Protein B modification: (SEP)357
Protein B sequence: PTKSPSAPIN
Protein B construct: PTKSP(pSer)APIN (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): 8.57
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: 4.64


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: MAP3K3
Protein B fragment: 352-361
Protein B site: 14-3-3_pSer357
Protein B modification: (SEP)357
Protein B sequence: PTKSPSAPIN
Protein B construct: PTKSP(pSer)APIN (crude synthetic peptide)

Average holdup BI: 0.84
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): 8.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.58


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: MAP3K3
Protein B fragment: 523-532
Protein B site: 14-3-3_pThr528
Protein B modification: (TPO)528
Protein B sequence: GMRSVTGTPY
Protein B construct: GMRSV(pThr)GTPY (crude synthetic peptide)

Average holdup BI: 0.77
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): 7.26
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: 4.42


Experiment series 5

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: MAP3K3
Protein B fragment: 352-361
Protein B site: 14-3-3_pSer357
Protein B modification: (SEP)357
Protein B sequence: PTKSPSAPIN
Protein B construct: PTKSP(pSer)APIN (crude synthetic peptide)

Average holdup BI: 0.78
Holdup BI standard deviation: 0.03
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): 7.05
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
Measured pKd: 4.51


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: MAP3K3
Protein B fragment: 289-298
Protein B site: 14-3-3_pThr294
Protein B modification: (TPO)294
Protein B sequence: SDGRRTFPRI
Protein B construct: SDGRR(pThr)FPRI (crude synthetic peptide)

Average holdup BI: 0.85
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): 7.02
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
Measured pKd: 4.63


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: MAP3K3
Protein B fragment: 289-298
Protein B site: 14-3-3_pThr294
Protein B modification: (TPO)294
Protein B sequence: SDGRRTFPRI
Protein B construct: SDGRR(pThr)FPRI (crude synthetic peptide)

Average holdup BI: 0.78
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): 6.06
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: 4.42


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: MAP3K3
Protein B fragment: 245-254
Protein B site: 14-3-3_pSer250
Protein B modification: (SEP)250
Protein B sequence: MSRAQSFPDN
Protein B construct: MSRAQ(pSer)FPDN (crude synthetic peptide)

Average holdup BI: 0.78
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): 5.88
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: 4.41


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: MAP3K3
Protein B fragment: 523-532
Protein B site: 14-3-3_pThr528
Protein B modification: (TPO)528
Protein B sequence: GMRSVTGTPY
Protein B construct: GMRSV(pThr)GTPY (crude synthetic peptide)

Average holdup BI: 0.82
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): 5.34
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: 4.54


Experiment series 10

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: MAP3K3
Protein B fragment: 523-532
Protein B site: 14-3-3_pThr528
Protein B modification: (TPO)528
Protein B sequence: GMRSVTGTPY
Protein B construct: GMRSV(pThr)GTPY (crude synthetic peptide)

Average holdup BI: 0.79
Holdup BI standard deviation: 0.11
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): 4.79
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: 4.52


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: MAP3K3
Protein B fragment: 245-254
Protein B site: 14-3-3_pSer250
Protein B modification: (SEP)250
Protein B sequence: MSRAQSFPDN
Protein B construct: MSRAQ(pSer)FPDN (crude synthetic peptide)

Average holdup BI: 0.66
Holdup BI standard deviation: 0.11
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): 4.43
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.25


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: MAP3K3
Protein B fragment: 289-298
Protein B site: 14-3-3_pThr294
Protein B modification: (TPO)294
Protein B sequence: SDGRRTFPRI
Protein B construct: SDGRR(pThr)FPRI (crude synthetic peptide)

Average holdup BI: 0.86
Holdup BI standard deviation: 0.1
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): 4.27
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: 4.74


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: MAP3K3
Protein B fragment: 161-170
Protein B site: 14-3-3_pSer166
Protein B modification: (SEP)166
Protein B sequence: RSRHLSVSSQ
Protein B construct: RSRHL(pSer)VSSQ (crude synthetic peptide)

Average holdup BI: 0.96
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): 3.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 5.27


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: MAP3K3
Protein B fragment: 335-344
Protein B site: 14-3-3_pSer340
Protein B modification: (SEP)340
Protein B sequence: LRSADSENAL
Protein B construct: LRSAD(pSer)ENAL (crude synthetic peptide)

Average holdup BI: 0.07
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): 3.31
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 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: MAP3K3
Protein B fragment: 332-341
Protein B site: 14-3-3_pSer337
Protein B modification: (SEP)337
Protein B sequence: RGRLRSADSE
Protein B construct: RGRLR(pSer)ADSE (crude synthetic peptide)

Average holdup BI: 0.47
Holdup BI standard deviation: 0.2
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): 1.67
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: 3.84


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: MAP3K3
Protein B fragment: 335-344
Protein B site: 14-3-3_pSer340
Protein B modification: (SEP)340
Protein B sequence: LRSADSENAL
Protein B construct: LRSAD(pSer)ENAL (crude synthetic peptide)

Average holdup BI: 0.04
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): 0.87
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 construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: MAP3K3
Protein B fragment: 511-520
Protein B site: 14-3-3_pThr516
Protein B modification: (TPO)516
Protein B sequence: SKRLQTICMS
Protein B construct: SKRLQ(pThr)ICMS (crude synthetic peptide)

Average holdup BI: 0.36
Holdup BI standard deviation: 0.5
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): 0.43
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 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: MAP3K3
Protein B fragment: 142-151
Protein B site: 14-3-3_pSer147
Protein B modification: (SEP)147
Protein B sequence: IKASQSAGDI
Protein B construct: IKASQ(pSer)AGDI (crude synthetic peptide)

Average holdup BI: 0.04
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): 0.35
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 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: MAP3K3
Protein B fragment: 142-151
Protein B site: 14-3-3_pSer147
Protein B modification: (SEP)147
Protein B sequence: IKASQSAGDI
Protein B construct: IKASQ(pSer)AGDI (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): 0.35
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: MAP3K3
Protein B fragment: 335-344
Protein B site: 14-3-3_pSer340
Protein B modification: (SEP)340
Protein B sequence: LRSADSENAL
Protein B construct: LRSAD(pSer)ENAL (crude synthetic peptide)

Average holdup BI: 0.02
Holdup BI standard deviation: 0.07
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 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: MAP3K3
Protein B fragment: 142-151
Protein B site: 14-3-3_pSer147
Protein B modification: (SEP)147
Protein B sequence: IKASQSAGDI
Protein B construct: IKASQ(pSer)AGDI (crude synthetic peptide)

Average holdup BI: 0.01
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): 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: MAP3K3
Protein B fragment: 312-321
Protein B site: 14-3-3_pThr317
Protein B modification: (TPO)317
Protein B sequence: SRSLSTNGEN
Protein B construct: SRSLS(pThr)NGEN (crude synthetic peptide)

Average holdup BI: -0.1
Holdup BI standard deviation: 0.91
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): 0.08
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.