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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...

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Autores principales: PANDEY, Kiran, NAHAR, Ashrafun, KADOKAWA, Hiroya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Japanese Society of Veterinary Science 2016
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.
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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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