Details of experiments between Q9BZF9 and P31946
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: UACA
Protein A fragment: 1348-1357
Protein A site: 14-3-3_pSer1353
Protein A modification: (SEP)1353
Protein A sequence: PTKRQSQLID
Protein A construct: PTKRQ(pSer)QLID (crude synthetic peptide)

Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta

Average holdup BI: -0.21
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): 0.8
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 2

Protein A protein: UACA
Protein A fragment: 1348-1357
Protein A site: 14-3-3_pSer1353
Protein A modification: (SEP)1353
Protein A sequence: PTKRQSQLID
Protein A construct: PTKRQ(pSer)QLID (crude synthetic peptide)

Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta

Average holdup BI: -0.09
Holdup BI standard deviation: 0.19
Immobilized partner concentration in holdup experiment (10-6M): 102
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: 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 3

Protein A protein: UACA
Protein A fragment: 621-630
Protein A site: 14-3-3_pSer626
Protein A modification: (SEP)626
Protein A sequence: AKLALSIPAE
Protein A construct: AKLAL(pSer)IPAE (crude synthetic peptide)

Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta

Average holdup BI: 0.08
Holdup BI standard deviation: 0.21
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.32
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 4

Protein A protein: UACA
Protein A fragment: 1043-1052
Protein A site: 14-3-3_pThr1048
Protein A modification: (TPO)1048
Protein A sequence: DLRDKTVLIE
Protein A construct: DLRDK(pThr)VLIE (crude synthetic peptide)

Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta

Average holdup BI: 0.02
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): 0.17
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 5

Protein A protein: UACA
Protein A fragment: 1043-1052
Protein A site: 14-3-3_pThr1048
Protein A modification: (TPO)1048
Protein A sequence: DLRDKTVLIE
Protein A construct: DLRDK(pThr)VLIE (crude synthetic peptide)

Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta

Average holdup BI: -0.01
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 102
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 6

Protein A protein: UACA
Protein A fragment: 621-630
Protein A site: 14-3-3_pSer626
Protein A modification: (SEP)626
Protein A sequence: AKLALSIPAE
Protein A construct: AKLAL(pSer)IPAE (crude synthetic peptide)

Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta

Average holdup BI: 0
Holdup BI standard deviation: 0.17
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): 0.01
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.