Minute™ ER(Endoplasmic reticulum) Enrichment Kit
Endoplasmic reticulum (ER) is a major membranous structure that functionally connects nuclear membrane
and plasma membrane. ER plays a central role in exocytic pathway of protein trafficking in all eukaryotic
cells. Proteins synthesized in cytoplasm are translocated into ER from where vesicles shuttle protein cargo
to the Golgi apparatus, which subsequently fuses with plasma membrane. Traditional methods for isolating
ER are based on density-gradient ultracentrifugation.The protocol requires a large amount of starting material
and the methods are tedious and time-consuming, and contain significant cross-contamination. Currently, all
commercial kits for ER isolation are based on methods developed in the 1970s. Unlike any other ER isolation
kit on the market, our ER isolation kit employs a patented spin-column-based technology that is simple and
rapid and requires only a small amount of starting cultured cells or tissues. This kit can differentially
precipitate native ER (mainlyrough ER) from cultured cells/tissues without using a Dounce homogenizer or
ultracentrifugation. The whole protocol can be done in about 2 hours.
Enrichment of ER from mouse tissue. A. Ponceau S stained blot. B. Western blot of ER, plasma membrance
and cytosolic markers. ER-hCalreticulin (R&D System, MAB38981). Na/K ATPase (Novus, NB3001 46SS);
GAPDH (Biolegend, 631401). 1. Total tissue lysate, 2 and 3. Extracted ER fraction in duplicate.