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Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation
Endoplasmic reticulum (ER) stress contributes to beta cell death in type 2 diabetes (T2DM). ER stress is characterized by increased level of ER stress markers such as C/EBP homologous protein (CHOP). Activation of CHOP leads to its translocation into the nucleus, where it induces cell death. We prev...
Autores principales: | , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Humana Press Inc
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2527405/ https://www.ncbi.nlm.nih.gov/pubmed/18651105 http://dx.doi.org/10.1007/s12013-008-9019-2 |
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author | Haataja, Leena Gurlo, Tatyana Huang, Chang-jiang Butler, Peter C. |
author_facet | Haataja, Leena Gurlo, Tatyana Huang, Chang-jiang Butler, Peter C. |
author_sort | Haataja, Leena |
collection | PubMed |
description | Endoplasmic reticulum (ER) stress contributes to beta cell death in type 2 diabetes (T2DM). ER stress is characterized by increased level of ER stress markers such as C/EBP homologous protein (CHOP). Activation of CHOP leads to its translocation into the nucleus, where it induces cell death. We previously reported nuclear CHOP in pancreatic sections from T2DM, but not T1DM, and in human islet amyloid polypeptide (IAPP) transgenic rodent pancreatic sections. These studies underscore the importance of studying nuclear CHOP. We have observed inconsistency in the detection of CHOP antibodies reported in the literature and also in our own experiments. To investigate the specificity of CHOP antibodies, we first induced ER stress by tunicamycin in rat insulinoma (INS) cells and prepared nuclear and cytoplasmic fractions. Then we examined CHOP expression by Western blotting and immunocytochemistry using seven commercially available CHOP antibodies in INS cells and human IAPP (h-IAPP) transgenic rodent pancreatic tissue. These studies show that three commercially available CHOP antibodies out of seven tested were non-specific. In conclusion, we give recommendations for CHOP antibody selection and methods to verify CHOP antibody specificity. Also, we propose that the authors report the catalog and lot numbers of the CHOP antibodies used. |
format | Text |
id | pubmed-2527405 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Humana Press Inc |
record_format | MEDLINE/PubMed |
spelling | pubmed-25274052008-09-03 Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation Haataja, Leena Gurlo, Tatyana Huang, Chang-jiang Butler, Peter C. Cell Biochem Biophys Technical Report Endoplasmic reticulum (ER) stress contributes to beta cell death in type 2 diabetes (T2DM). ER stress is characterized by increased level of ER stress markers such as C/EBP homologous protein (CHOP). Activation of CHOP leads to its translocation into the nucleus, where it induces cell death. We previously reported nuclear CHOP in pancreatic sections from T2DM, but not T1DM, and in human islet amyloid polypeptide (IAPP) transgenic rodent pancreatic sections. These studies underscore the importance of studying nuclear CHOP. We have observed inconsistency in the detection of CHOP antibodies reported in the literature and also in our own experiments. To investigate the specificity of CHOP antibodies, we first induced ER stress by tunicamycin in rat insulinoma (INS) cells and prepared nuclear and cytoplasmic fractions. Then we examined CHOP expression by Western blotting and immunocytochemistry using seven commercially available CHOP antibodies in INS cells and human IAPP (h-IAPP) transgenic rodent pancreatic tissue. These studies show that three commercially available CHOP antibodies out of seven tested were non-specific. In conclusion, we give recommendations for CHOP antibody selection and methods to verify CHOP antibody specificity. Also, we propose that the authors report the catalog and lot numbers of the CHOP antibodies used. Humana Press Inc 2008-07-24 2008-07 /pmc/articles/PMC2527405/ /pubmed/18651105 http://dx.doi.org/10.1007/s12013-008-9019-2 Text en © Humana Press 2008 |
spellingShingle | Technical Report Haataja, Leena Gurlo, Tatyana Huang, Chang-jiang Butler, Peter C. Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation |
title | Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation |
title_full | Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation |
title_fullStr | Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation |
title_full_unstemmed | Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation |
title_short | Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation |
title_sort | many commercially available antibodies for detection of chop expression as a marker of endoplasmic reticulum stress fail specificity evaluation |
topic | Technical Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2527405/ https://www.ncbi.nlm.nih.gov/pubmed/18651105 http://dx.doi.org/10.1007/s12013-008-9019-2 |
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