Preparation and extraction of chorion proteins from Salmo salar embryos at the pigmented eye stage for electrophoresis with SDS-polyacrylamide gel
The chorion fulfills important functions in fish embryos, including protecting the embryo during development. The characterization of the protein profile of this envelope could be used as a bioindicator in the evaluation of the quality of embryonic development. The object of this work was to validat...
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ftdoajarticles:oai:doaj.org/article:47915a403bbf4b35bcab6bc01c01fb28 2024-01-28T10:04:32+01:00 Preparation and extraction of chorion proteins from Salmo salar embryos at the pigmented eye stage for electrophoresis with SDS-polyacrylamide gel Maritza Pérez-Atehortúa Stefania E. Short Cristian Aranzaez-Rios Jorge Farías Ricardo Pinheiro S. Oliveira Wellison Amorim Pereira Jennie Risopatrón Iván Valdebenito Elías Figueroa Villalobos 2024-06-01T00:00:00Z https://doi.org/10.1016/j.mex.2023.102533 https://doaj.org/article/47915a403bbf4b35bcab6bc01c01fb28 EN eng Elsevier http://www.sciencedirect.com/science/article/pii/S2215016123005290 https://doaj.org/toc/2215-0161 2215-0161 doi:10.1016/j.mex.2023.102533 https://doaj.org/article/47915a403bbf4b35bcab6bc01c01fb28 MethodsX, Vol 12, Iss , Pp 102533- (2024) Method of preparation and extraction of chorion proteins from embryos at 280 accumulated thermal units (ATU) of Atlantic salmon (Salmo salar) for protein characterization by SDS-PAGE electrophoresis Science Q article 2024 ftdoajarticles https://doi.org/10.1016/j.mex.2023.102533 2023-12-31T01:41:19Z The chorion fulfills important functions in fish embryos, including protecting the embryo during development. The characterization of the protein profile of this envelope could be used as a bioindicator in the evaluation of the quality of embryonic development. The object of this work was to validate a standardized protocol for protein extraction from chorion of Salmo salar embryos at 280 accumulated thermal units (ATU) by comparing and combining existing methods. The protocol consists of consecutive washing of the chorion samples followed by protein extraction with the solution that was named SDS solution (Tris–HCl 100 mM (pH 8), Urea 8 M, 1% SDS, β-mercaptoethanol 300 mM and EGTA 10 Mm, and 1% protease inhibitor cocktail) and mechanical methods. Protein extraction is enhanced by a working temperature of 75 °C and use of a disperser. The protein concentration was quantified by Bradford Assay. After extraction, the samples were diluted (dilution factor 10) before reading against the calibration curve. After gel electrophoresis with a load of 3 µg of protein, staining showed more than 4 bands, with molecular weights between 25 kDa and 180 kDa. • The protein profile of fish chorion was between 25 kDa and 180 kDa. • Solution containing 1% SDS allows a higher extraction of proteins from the chorion of Atlantic salmon embryos with 280 ATU. • Chorion protein identification is a valuable tool in determining gamete and embryo quality in fish. Article in Journal/Newspaper Atlantic salmon Salmo salar Directory of Open Access Journals: DOAJ Articles MethodsX 12 102533 |
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Directory of Open Access Journals: DOAJ Articles |
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language |
English |
topic |
Method of preparation and extraction of chorion proteins from embryos at 280 accumulated thermal units (ATU) of Atlantic salmon (Salmo salar) for protein characterization by SDS-PAGE electrophoresis Science Q |
spellingShingle |
Method of preparation and extraction of chorion proteins from embryos at 280 accumulated thermal units (ATU) of Atlantic salmon (Salmo salar) for protein characterization by SDS-PAGE electrophoresis Science Q Maritza Pérez-Atehortúa Stefania E. Short Cristian Aranzaez-Rios Jorge Farías Ricardo Pinheiro S. Oliveira Wellison Amorim Pereira Jennie Risopatrón Iván Valdebenito Elías Figueroa Villalobos Preparation and extraction of chorion proteins from Salmo salar embryos at the pigmented eye stage for electrophoresis with SDS-polyacrylamide gel |
topic_facet |
Method of preparation and extraction of chorion proteins from embryos at 280 accumulated thermal units (ATU) of Atlantic salmon (Salmo salar) for protein characterization by SDS-PAGE electrophoresis Science Q |
description |
The chorion fulfills important functions in fish embryos, including protecting the embryo during development. The characterization of the protein profile of this envelope could be used as a bioindicator in the evaluation of the quality of embryonic development. The object of this work was to validate a standardized protocol for protein extraction from chorion of Salmo salar embryos at 280 accumulated thermal units (ATU) by comparing and combining existing methods. The protocol consists of consecutive washing of the chorion samples followed by protein extraction with the solution that was named SDS solution (Tris–HCl 100 mM (pH 8), Urea 8 M, 1% SDS, β-mercaptoethanol 300 mM and EGTA 10 Mm, and 1% protease inhibitor cocktail) and mechanical methods. Protein extraction is enhanced by a working temperature of 75 °C and use of a disperser. The protein concentration was quantified by Bradford Assay. After extraction, the samples were diluted (dilution factor 10) before reading against the calibration curve. After gel electrophoresis with a load of 3 µg of protein, staining showed more than 4 bands, with molecular weights between 25 kDa and 180 kDa. • The protein profile of fish chorion was between 25 kDa and 180 kDa. • Solution containing 1% SDS allows a higher extraction of proteins from the chorion of Atlantic salmon embryos with 280 ATU. • Chorion protein identification is a valuable tool in determining gamete and embryo quality in fish. |
format |
Article in Journal/Newspaper |
author |
Maritza Pérez-Atehortúa Stefania E. Short Cristian Aranzaez-Rios Jorge Farías Ricardo Pinheiro S. Oliveira Wellison Amorim Pereira Jennie Risopatrón Iván Valdebenito Elías Figueroa Villalobos |
author_facet |
Maritza Pérez-Atehortúa Stefania E. Short Cristian Aranzaez-Rios Jorge Farías Ricardo Pinheiro S. Oliveira Wellison Amorim Pereira Jennie Risopatrón Iván Valdebenito Elías Figueroa Villalobos |
author_sort |
Maritza Pérez-Atehortúa |
title |
Preparation and extraction of chorion proteins from Salmo salar embryos at the pigmented eye stage for electrophoresis with SDS-polyacrylamide gel |
title_short |
Preparation and extraction of chorion proteins from Salmo salar embryos at the pigmented eye stage for electrophoresis with SDS-polyacrylamide gel |
title_full |
Preparation and extraction of chorion proteins from Salmo salar embryos at the pigmented eye stage for electrophoresis with SDS-polyacrylamide gel |
title_fullStr |
Preparation and extraction of chorion proteins from Salmo salar embryos at the pigmented eye stage for electrophoresis with SDS-polyacrylamide gel |
title_full_unstemmed |
Preparation and extraction of chorion proteins from Salmo salar embryos at the pigmented eye stage for electrophoresis with SDS-polyacrylamide gel |
title_sort |
preparation and extraction of chorion proteins from salmo salar embryos at the pigmented eye stage for electrophoresis with sds-polyacrylamide gel |
publisher |
Elsevier |
publishDate |
2024 |
url |
https://doi.org/10.1016/j.mex.2023.102533 https://doaj.org/article/47915a403bbf4b35bcab6bc01c01fb28 |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_source |
MethodsX, Vol 12, Iss , Pp 102533- (2024) |
op_relation |
http://www.sciencedirect.com/science/article/pii/S2215016123005290 https://doaj.org/toc/2215-0161 2215-0161 doi:10.1016/j.mex.2023.102533 https://doaj.org/article/47915a403bbf4b35bcab6bc01c01fb28 |
op_doi |
https://doi.org/10.1016/j.mex.2023.102533 |
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MethodsX |
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12 |
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102533 |
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1789330277220220928 |