Details of experiments between P61981 and Q5JSL3
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: DOCK11
Protein B fragment: 1347-1356
Protein B site: 14-3-3_pThr1352
Protein B modification: (TPO)1352
Protein B sequence: DRKSQTMPAL
Protein B construct: DRKSQ(pThr)MPAL (crude synthetic peptide)

Measured dissociation constant (10-6M): <1
Immobilized partner concentration in holdup experiment (10-6M): 129
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: 1
Measured pKd: 6


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: DOCK11
Protein B fragment: 53-62
Protein B site: 14-3-3_pSer58
Protein B modification: (SEP)58
Protein B sequence: KTQIYSDPLR
Protein B construct: KTQIY(pSer)DPLR (crude synthetic peptide)

Average holdup BI: 0.93
Holdup BI standard deviation: 0
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): 10.66
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: 4.98


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: DOCK11
Protein B fragment: 53-62
Protein B site: 14-3-3_pSer58
Protein B modification: (SEP)58
Protein B sequence: KTQIYSDPLR
Protein B construct: KTQIY(pSer)DPLR (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): 7.78
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: 5.02


Experiment series 4

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: DOCK11
Protein B fragment: 53-62
Protein B site: 14-3-3_pSer58
Protein B modification: (SEP)58
Protein B sequence: KTQIYSDPLR
Protein B construct: KTQIY(pSer)DPLR (crude synthetic peptide)

Average holdup BI: 0.9
Holdup BI standard deviation: 0.01
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): 7.2
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
Measured pKd: 4.9


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: DOCK11
Protein B fragment: 1347-1356
Protein B site: 14-3-3_pThr1352
Protein B modification: (TPO)1352
Protein B sequence: DRKSQTMPAL
Protein B construct: DRKSQ(pThr)MPAL (crude synthetic peptide)

Average holdup BI: 0.97
Holdup BI standard deviation: 0
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): 5.41
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: 5.35


Experiment series 6

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: DOCK11
Protein B fragment: 82-91
Protein B site: 14-3-3_pThr87
Protein B modification: (TPO)87
Protein B sequence: RTVQSTVPED
Protein B construct: RTVQS(pThr)VPED (crude synthetic peptide)

Average holdup BI: 0.17
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): 5.14
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 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: DOCK11
Protein B fragment: 495-504
Protein B site: 14-3-3_pSer500
Protein B modification: (SEP)500
Protein B sequence: PYIKNSDPVK
Protein B construct: PYIKN(pSer)DPVK (crude synthetic peptide)

Average holdup BI: 0.15
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): 2.95
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 8

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: DOCK11
Protein B fragment: 1232-1241
Protein B site: 14-3-3_pSer1237
Protein B modification: (SEP)1237
Protein B sequence: IKREDSRGSL
Protein B construct: IKRED(pSer)RGSL (crude synthetic peptide)

Average holdup BI: 0.53
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): 2.23
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


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: DOCK11
Protein B fragment: 82-91
Protein B site: 14-3-3_pThr87
Protein B modification: (TPO)87
Protein B sequence: RTVQSTVPED
Protein B construct: RTVQS(pThr)VPED (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): 1.47
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 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: DOCK11
Protein B fragment: 1232-1241
Protein B site: 14-3-3_pSer1237
Protein B modification: (SEP)1237
Protein B sequence: IKREDSRGSL
Protein B construct: IKRED(pSer)RGSL (crude synthetic peptide)

Average holdup BI: 0.14
Holdup BI standard deviation: 0.12
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): 1.14
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 11

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: DOCK11
Protein B fragment: 7-16
Protein B site: 14-3-3_pSer12
Protein B modification: (SEP)12
Protein B sequence: FTKRLSKPGT
Protein B construct: FTKRL(pSer)KPGT (crude synthetic peptide)

Average holdup BI: 0.07
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): 1.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 12

Protein A protein: 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: DOCK11
Protein B fragment: 82-91
Protein B site: 14-3-3_pThr87
Protein B modification: (TPO)87
Protein B sequence: RTVQSTVPED
Protein B construct: RTVQS(pThr)VPED (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): 0.83
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 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: DOCK11
Protein B fragment: 495-504
Protein B site: 14-3-3_pSer500
Protein B modification: (SEP)500
Protein B sequence: PYIKNSDPVK
Protein B construct: PYIKN(pSer)DPVK (crude synthetic peptide)

Average holdup BI: 0.09
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): 0.8
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
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B protein: DOCK11
Protein B fragment: 7-16
Protein B site: 14-3-3_pSer12
Protein B modification: (SEP)12
Protein B sequence: FTKRLSKPGT
Protein B construct: FTKRL(pSer)KPGT (crude synthetic peptide)

Average holdup BI: 0.03
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): 0.46
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: DOCK11
Protein B fragment: 324-333
Protein B site: 14-3-3_pSer329
Protein B modification: (SEP)329
Protein B sequence: QLNKLSRGDG
Protein B construct: QLNKL(pSer)RGDG (crude synthetic peptide)

Average holdup BI: 0.1
Holdup BI standard deviation: 0.27
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): 0.31
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: 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: DOCK11
Protein B fragment: 495-504
Protein B site: 14-3-3_pSer500
Protein B modification: (SEP)500
Protein B sequence: PYIKNSDPVK
Protein B construct: PYIKN(pSer)DPVK (crude synthetic peptide)

Average holdup BI: 0.03
Holdup BI standard deviation: 0.11
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): 0.21
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 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: DOCK11
Protein B fragment: 324-333
Protein B site: 14-3-3_pSer329
Protein B modification: (SEP)329
Protein B sequence: QLNKLSRGDG
Protein B construct: QLNKL(pSer)RGDG (crude synthetic peptide)

Average holdup BI: -0.06
Holdup BI standard deviation: 0.25
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): 0.18
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 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: DOCK11
Protein B fragment: 7-16
Protein B site: 14-3-3_pSer12
Protein B modification: (SEP)12
Protein B sequence: FTKRLSKPGT
Protein B construct: FTKRL(pSer)KPGT (crude synthetic peptide)

Average holdup BI: 0.02
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): 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 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: DOCK11
Protein B fragment: 324-333
Protein B site: 14-3-3_pSer329
Protein B modification: (SEP)329
Protein B sequence: QLNKLSRGDG
Protein B construct: QLNKL(pSer)RGDG (crude synthetic peptide)

Average holdup BI: 0.02
Holdup BI standard deviation: 0.23
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.