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Method for isolating pure bovine gonadotrophs from anterior pituitary using magnetic nanoparticles and anti-gonadotropin-releasing hormone receptor antibody
No methods are currently available for rapidly isolating gonadotrophs from the anterior pituitary (AP) in any species. We developed a method for preparing pure bovine gonadotrophs from a heterogeneous AP cell mixture by magnetic separation and our original antibody against the N terminus of bovine g...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Japanese Society of Veterinary Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5138424/ https://www.ncbi.nlm.nih.gov/pubmed/27430292 http://dx.doi.org/10.1292/jvms.16-0157 |
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author | PANDEY, Kiran NAHAR, Ashrafun KADOKAWA, Hiroya |
author_facet | PANDEY, Kiran NAHAR, Ashrafun KADOKAWA, Hiroya |
author_sort | PANDEY, Kiran |
collection | PubMed |
description | No methods are currently available for rapidly isolating gonadotrophs from the anterior pituitary (AP) in any species. We developed a method for preparing pure bovine gonadotrophs from a heterogeneous AP cell mixture by magnetic separation and our original antibody against the N terminus of bovine gonadotropin-releasing hormone receptor (GnRHR). A bovine AP cell mixture was incubated with the anti-GnRHR antibody, anti-dextran antibody-conjugated secondary antibody and dextran-coated magnetic nanoparticles for magnetic isolation. Approximately 5.2 × 10(6) cells were isolated per AP of Japanese Black heifers (26 months of age) and cultured, and confocal microscopy confirmed to be GnRHR- and luteinizing hormone-positive, corresponding to a purity of 100%. Approximately 44.5 µg of total protein was extracted from the pure gonadotrophs per AP. |
format | Online Article Text |
id | pubmed-5138424 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | The Japanese Society of Veterinary Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-51384242016-12-06 Method for isolating pure bovine gonadotrophs from anterior pituitary using magnetic nanoparticles and anti-gonadotropin-releasing hormone receptor antibody PANDEY, Kiran NAHAR, Ashrafun KADOKAWA, Hiroya J Vet Med Sci Theriogenology No methods are currently available for rapidly isolating gonadotrophs from the anterior pituitary (AP) in any species. We developed a method for preparing pure bovine gonadotrophs from a heterogeneous AP cell mixture by magnetic separation and our original antibody against the N terminus of bovine gonadotropin-releasing hormone receptor (GnRHR). A bovine AP cell mixture was incubated with the anti-GnRHR antibody, anti-dextran antibody-conjugated secondary antibody and dextran-coated magnetic nanoparticles for magnetic isolation. Approximately 5.2 × 10(6) cells were isolated per AP of Japanese Black heifers (26 months of age) and cultured, and confocal microscopy confirmed to be GnRHR- and luteinizing hormone-positive, corresponding to a purity of 100%. Approximately 44.5 µg of total protein was extracted from the pure gonadotrophs per AP. The Japanese Society of Veterinary Science 2016-07-16 2016-11 /pmc/articles/PMC5138424/ /pubmed/27430292 http://dx.doi.org/10.1292/jvms.16-0157 Text en ©2016 The Japanese Society of Veterinary Science http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. |
spellingShingle | Theriogenology PANDEY, Kiran NAHAR, Ashrafun KADOKAWA, Hiroya Method for isolating pure bovine gonadotrophs from anterior pituitary using magnetic nanoparticles and anti-gonadotropin-releasing hormone receptor antibody |
title | Method for isolating pure bovine gonadotrophs from anterior pituitary using
magnetic nanoparticles and anti-gonadotropin-releasing hormone receptor
antibody |
title_full | Method for isolating pure bovine gonadotrophs from anterior pituitary using
magnetic nanoparticles and anti-gonadotropin-releasing hormone receptor
antibody |
title_fullStr | Method for isolating pure bovine gonadotrophs from anterior pituitary using
magnetic nanoparticles and anti-gonadotropin-releasing hormone receptor
antibody |
title_full_unstemmed | Method for isolating pure bovine gonadotrophs from anterior pituitary using
magnetic nanoparticles and anti-gonadotropin-releasing hormone receptor
antibody |
title_short | Method for isolating pure bovine gonadotrophs from anterior pituitary using
magnetic nanoparticles and anti-gonadotropin-releasing hormone receptor
antibody |
title_sort | method for isolating pure bovine gonadotrophs from anterior pituitary using
magnetic nanoparticles and anti-gonadotropin-releasing hormone receptor
antibody |
topic | Theriogenology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5138424/ https://www.ncbi.nlm.nih.gov/pubmed/27430292 http://dx.doi.org/10.1292/jvms.16-0157 |
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