Details of experiments between P61981 and Q8WUI4
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: HDAC7
Protein B fragment: 481-490
Protein B site: 14-3-3_pSer486
Protein B modification: (SEP)486
Protein B sequence: LSRAQSSPAA
Protein B construct: LSRAQ(pSer)SPAA (crude synthetic peptide)

Average holdup BI: 0.98
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): 10.63
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: 5.65


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: HDAC7
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: LRKTVSEPNL
Protein B construct: LRKTV(pSer)EPNL (crude synthetic peptide)

Average holdup BI: 0.97
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): 9.03
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
Measured pKd: 5.47


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: HDAC7
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: LRKTVSEPNL
Protein B construct: LRKTV(pSer)EPNL (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): 8.41
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: 5.43


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: HDAC7
Protein B fragment: 150-159
Protein B site: 14-3-3_pSer155
Protein B modification: (SEP)155
Protein B sequence: LRKTVSEPNL
Protein B construct: LRKTV(pSer)EPNL (crude synthetic peptide)

Average holdup BI: 0.95
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): 8.35
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
Measured pKd: 5.13


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: HDAC7
Protein B fragment: 176-185
Protein B site: 14-3-3_pSer181
Protein B modification: (SEP)181
Protein B sequence: LLRKESAPPS
Protein B construct: LLRKE(pSer)APPS (crude synthetic peptide)

Average holdup BI: 0.37
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): 7.76
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: 3.67


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: HDAC7
Protein B fragment: 481-490
Protein B site: 14-3-3_pSer486
Protein B modification: (SEP)486
Protein B sequence: LSRAQSSPAA
Protein B construct: LSRAQ(pSer)SPAA (crude synthetic peptide)

Average holdup BI: 0.98
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): 7.39
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: 5.62


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: HDAC7
Protein B fragment: 353-362
Protein B site: 14-3-3_pSer358
Protein B modification: (SEP)358
Protein B sequence: LSRTRSEPLP
Protein B construct: LSRTR(pSer)EPLP (crude synthetic peptide)

Average holdup BI: 0.8
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): 6.6
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: 4.56


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: HDAC7
Protein B fragment: 481-490
Protein B site: 14-3-3_pSer486
Protein B modification: (SEP)486
Protein B sequence: LSRAQSSPAA
Protein B construct: LSRAQ(pSer)SPAA (crude synthetic peptide)

Average holdup BI: 0.97
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): 5.83
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: 5.5


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: HDAC7
Protein B fragment: 353-362
Protein B site: 14-3-3_pSer358
Protein B modification: (SEP)358
Protein B sequence: LSRTRSEPLP
Protein B construct: LSRTR(pSer)EPLP (crude synthetic peptide)

Average holdup BI: 0.89
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): 5.56
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
Measured pKd: 4.77


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: HDAC7
Protein B fragment: 176-185
Protein B site: 14-3-3_pSer181
Protein B modification: (SEP)181
Protein B sequence: LLRKESAPPS
Protein B construct: LLRKE(pSer)APPS (crude synthetic peptide)

Average holdup BI: 0.34
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): 4.97
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: HDAC7
Protein B fragment: 353-362
Protein B site: 14-3-3_pSer358
Protein B modification: (SEP)358
Protein B sequence: LSRTRSEPLP
Protein B construct: LSRTR(pSer)EPLP (crude synthetic peptide)

Average holdup BI: 0.69
Holdup BI standard deviation: 0.16
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): 3.78
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: 4.23


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: HDAC7
Protein B fragment: 400-409
Protein B site: 14-3-3_pSer405
Protein B modification: (SEP)405
Protein B sequence: LRQIPSAEDL
Protein B construct: LRQIP(pSer)AEDL (crude synthetic peptide)

Average holdup BI: -0.19
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): 2.53
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: 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: HDAC7
Protein B fragment: 176-185
Protein B site: 14-3-3_pSer181
Protein B modification: (SEP)181
Protein B sequence: LLRKESAPPS
Protein B construct: LLRKE(pSer)APPS (crude synthetic peptide)

Average holdup BI: 0.22
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): 1.72
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: HDAC7
Protein B fragment: 502-511
Protein B site: 14-3-3_pSer507
Protein B modification: (SEP)507
Protein B sequence: QARVLSSSET
Protein B construct: QARVL(pSer)SSET (crude synthetic peptide)

Average holdup BI: 0.03
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): 0.53
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: HDAC7
Protein B fragment: 502-511
Protein B site: 14-3-3_pSer507
Protein B modification: (SEP)507
Protein B sequence: QARVLSSSET
Protein B construct: QARVL(pSer)SSET (crude synthetic peptide)

Average holdup BI: 0.02
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): 0.42
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 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: HDAC7
Protein B fragment: 400-409
Protein B site: 14-3-3_pSer405
Protein B modification: (SEP)405
Protein B sequence: LRQIPSAEDL
Protein B construct: LRQIP(pSer)AEDL (crude synthetic peptide)

Average holdup BI: 0.01
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): 0.41
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
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: HDAC7
Protein B fragment: 459-468
Protein B site: 14-3-3_pSer464
Protein B modification: (SEP)464
Protein B sequence: RLPRGSTGDT
Protein B construct: RLPRG(pSer)TGDT (crude synthetic peptide)

Average holdup BI: 0.11
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): 0.35
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 18

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: HDAC7
Protein B fragment: 459-468
Protein B site: 14-3-3_pSer464
Protein B modification: (SEP)464
Protein B sequence: RLPRGSTGDT
Protein B construct: RLPRG(pSer)TGDT (crude synthetic peptide)

Average holdup BI: -0.08
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): 0.26
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: HDAC7
Protein B fragment: 459-468
Protein B site: 14-3-3_pSer464
Protein B modification: (SEP)464
Protein B sequence: RLPRGSTGDT
Protein B construct: RLPRG(pSer)TGDT (crude synthetic peptide)

Average holdup BI: 0.02
Holdup BI standard deviation: 0.1
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 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: HDAC7
Protein B fragment: 502-511
Protein B site: 14-3-3_pSer507
Protein B modification: (SEP)507
Protein B sequence: QARVLSSSET
Protein B construct: QARVL(pSer)SSET (crude synthetic peptide)

Average holdup BI: 0.01
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): 0.18
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 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: HDAC7
Protein B fragment: 400-409
Protein B site: 14-3-3_pSer405
Protein B modification: (SEP)405
Protein B sequence: LRQIPSAEDL
Protein B construct: LRQIP(pSer)AEDL (crude synthetic peptide)

Average holdup BI: -0.03
Holdup BI standard deviation: 0.14
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.