Abstract
The method described here is intended to study intracellular localization of proteins in colon cells. This protocol was used to localize IRE1β in the endoplasmic reticulum membrane. We used anti-IRE1β antibody raised in guinea pig for this purpose. We also studied the location of BiP (also known as GRP78), with the antibody raised in rabbit. Both antibodies used with appropriate gold particle-conjugated secondary antibodies gave good results. Primary mouse antibodies are not recommended because secondary anti-mouse antibodies also react with the internal mouse epitopes.
Keywords: Immunoelectron microscopy, Goblet cells, ER stress, IRE1, BiP
Materials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
This work was supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI Grants 24228002, 24248019 (to K.K.), and 14580699 (to A.T.), Ministry of Education, Culture, Sports, Science and Technology in Japan (MEXT) KAKENHI Grant 19058010 (to K.K.), The Uehara Memorial Foundation (K.K.), Takeda Science Foundation (K.K.), Mitsubishi Foundation (K.K.).
References
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