Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation
Open Access
- 24 July 2008
- journal article
- research article
- Published by Springer Science and Business Media LLC in Cell Biochemistry and Biophysics
- Vol. 51 (2-3), 105-107
- https://doi.org/10.1007/s12013-008-9019-2
Abstract
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.Keywords
This publication has 6 references indexed in Scilit:
- Induction of endoplasmic reticulum stress-induced β-cell apoptosis and accumulation of polyubiquitinated proteins by human islet amyloid polypeptideAmerican Journal of Physiology-Endocrinology and Metabolism, 2007
- The endoplasmic reticulum in pancreatic beta cells of type 2 diabetes patientsDiabetologia, 2007
- High Expression Rates of Human Islet Amyloid Polypeptide Induce Endoplasmic Reticulum Stress–Mediated β-Cell Apoptosis, a Characteristic of Humans With Type 2 but Not Type 1 DiabetesDiabetes, 2007
- Endoplasmic reticulum stress contributes to beta cell apoptosis in type 2 diabetesDiabetologia, 2007
- Endoplasmic Reticulum Stress Links Obesity, Insulin Action, and Type 2 DiabetesScience, 2004
- Roles of CHOP/GADD153 in endoplasmic reticulum stressCell Death & Differentiation, 2003