Dataset for "Membrane extraction with styrene-maleic acid copolymer results in insulin receptor autophosphorylation in the absence of ligand"

The data was collected to establish whether the insulin receptor maintained its native functionality once extracted into polymer nanodiscs using styrene maleic acid (SMA). The data gathered here show the raw .tiff files of the western blots. Differences in protein activity were quantified by comparing changes between total and phospho- antibody detection, using densitometry. Downstream subunits associated with the insulin receptor cascade were also quantified in this way.

.zip file containing the following items;
1. all raw .tiff files gathered from western blots used for quantification. A MS excel spreadsheet is also included to indicate which bands from the experiments were used for each quantification.
2. all raw .tiff files used for representative blots in each figure, with MS Powerpoint document to show annoted versions for these images.
3. READ_ME.txt file to further describe the data present.
4. Excel spreadsheet with raw data values from densitometry analysis from western blots, representative western blots for each experiment, calculations from the densitometry values and statistics associated with these values.

Subjects:
Biomolecules and biochemistry

Cite this dataset as:
Morrison, K., Whitley, P., Koumanov, F., 2022. Dataset for "Membrane extraction with styrene-maleic acid copolymer results in insulin receptor autophosphorylation in the absence of ligand". Bath: University of Bath Research Data Archive. Available from: https://doi.org/10.15125/BATH-01041.

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Data

For_archive.zip
application/zip (192MB)
Creative Commons: Attribution 4.0

.zip file containing the following items for the paper; 1. all raw .tiff files for western blots used for quantification including MS excel spreadsheet depicting which bands from the experiments were used. 2. all raw .tiff files used for representative blots in each figure, with MS Powerpoint document to show annoted versions. 3. READ_ME.txt file to further describe the data present. 4. Denistometry values, calculations and statistics; collated_InsR_values_for_archive.xlsx

Aimed at potential readers of journal manuscript "Membrane Extraction with Styrene-Maleic Acid Copolymer Results in Insulin Receptor Autophosphorylation in the Absence of Ligand."

Creators

Paul Whitley
University of Bath

Contributors

University of Bath
Rights Holder

Documentation

Data collection method:

Samples were generated using methods discussed in paper manuscript "Membrane Extraction with Styrene-Maleic Acid Copolymer Results in Insulin Receptor Autophosphorylation in the Absence of Ligand." Samples were separated on 10% SDS polyacrylamide gels and transferred onto nitrocellulose using a semi-dry transfer apparatus, before immunodetection with primary antibodies; either total antibodies or phopho-specific antibodies. Blots were incubated with secondary antibody,HRP, followed by washes. ECL™ Select Western blotting detection reagent was used for detection. Images were acquired with EPI Chemi II darkroom (UVP).

Technical details and requirements:

Quantification of bands on immunoblots was measured by densitometry using Image Studio Lite (LI-COR Biosciences®). Specific band intensities were normalized to total Akt levels for that sample. The ratios between phospho-specific and total protein band intensities were calculated in the same sample. Data has been deposited and can be opened with Microsoft Excel.

Additional information:

A READ_ME.txt file is included to provide further information of the data present in the .zip file. Folders labelled "Figure_X", where X corresponds to the figure number, contain raw .tiff files used for representative blots in the paper. An additional MS PowerPoint file is included to annotate these images. "SX" describes the supplementary figures. All_blots_for_quantification folder contains all raw .tiff files obtained and included in quantification of densitometry values. Experiments_used_for_each_graph.xlsx highlights the experiments and bands used for each quantification, representated by graphs in the paper. The Excel spreadsheet "collated_InsR_values_for_archive.xlsx" has three tabs; 1. Densitometry_values - holds the raw densitometry values calculated with Image Studio Lite (LI-COR Biosciences®); normalisation to Akt in separate column (L); representative western blots for each sample measured and "fold difference" between basal and + Insulin samples. 2. Calculations - taking the fold difference tables for each experiment, with additional tables calculating fold difference relative to POST SMA Basal (Green) and PRE RIPA Basal (Orange). Averages were taken for each sample across multiple experiments and standard deviation (SD) and standard error mean (SEM) were caculated. 3. Stats - Statistic for each protein of study; InsR, IRS and Akt, across multiple experiments using paired t-tests.

Documentation Files

READ_ME.txt
text/plain (1kB)
Creative Commons: Attribution 4.0

Funders

Engineering and Physical Sciences Research Council (EPSRC)
https://doi.org/10.13039/501100000266

EPSRC Doctoral Training Centre in Sustainable Chemical Technologies
EP/L016354/1

Science and Technology Facilities Council (STFC)
https://doi.org/10.13039/501100000271

Grant
SA-54

Medical Research Council (MRC)
https://doi.org/10.13039/501100000265

Grant
MR/P0002927/1

Grant
19/0005983

Publication details

Publication date: 3 March 2022
by: University of Bath

Version: 1

DOI: https://doi.org/10.15125/BATH-01041

URL for this record: https://researchdata.bath.ac.uk/id/eprint/1041

Related papers and books

Morrison, K. A., Wood, L., Edler, K. J., Doutch, J., Price, G. J., Koumanov, F., and Whitley, P., 2022. Membrane extraction with styrene-maleic acid copolymer results in insulin receptor autophosphorylation in the absence of ligand. Scientific Reports, 12(1). Available from: https://doi.org/10.1038/s41598-022-07606-5.

Contact information

Please contact the Research Data Service in the first instance for all matters concerning this item.

Contact person: Paul Whitley

Departments:

Faculty of Humanities & Social Sciences
Health

Life Sciences
Biology & Biochemistry

Faculty of Science
Chemistry

Research Centres & Institutes
Centre for Sustainable Chemical Technologies