Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation

Authors: Haataja, Leena¹; Gurlo, Tatyana¹; Huang, Chang-jiang¹; Butler, Peter²

Source: Cell Biochemistry and Biophysics, Volume 51, Numbers 2-3, July 2008 , pp. 105-107(3)

Buy & download fulltext article:

Price: $42.00 plus tax (Refund Policy)

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: Endoplasmic reticulum stress; CHOP; Diabetes; Islet amyloid polypeptide

Document Type: Research article

DOI: http://dx.doi.org/10.1007/s12013-008-9019-2

Affiliations: 1: Larry Hillblom Islet Research Center, UCLA David Geffen School of Medicine, 900 Weyburn Place #A, Los Angeles, CA, 90024-2852, USA, 2: Larry Hillblom Islet Research Center, UCLA David Geffen School of Medicine, 900 Weyburn Place #A, Los Angeles, CA, 90024-2852, USA, Email: pbutler@mednet.ucla.edu

Publication date: 2008-07-01

Many Commercially Available Antibodies for Detection of CHOP Expression as a Marker of Endoplasmic Reticulum Stress Fail Specificity Evaluation

Buy & download fulltext article:

Tools

Key

Text size:

Useful pages