Organelles: Endoplasmic Reticulum
Overview
The endoplasmic reticulum (ER) is a network of tubular membrane-enclosed spaces that contain enzymes responsible for lipid biosynthesis and the early stages of protein post-translational modification. The cytoplasmic face of the rough ER membrane is studded with ribosomes that inject elongating nascent proteins into the ER.
Data Examples
Protein Disulfide Isomerase/P4HB in HeLa Human Cell Line. Protein Disulfide Isomerase/P4HB was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human Protein Disulfide Isomerase/P4HB Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF4236) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Calreticulin in HeLa Human Cell Line. Calreticulin was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human Calreticulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3898) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to endoplasmic reticula. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human and Rat GRP78/HSPA5 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and A20 mouse B cell lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Human/Mouse/Rat GRP78/HSPA5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4846) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for GRP78/HSPA5 at approximately 80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Visualization of the Endoplasmic Reticulum in PNT2 Cells. ReZolve-ER™ (20 mM; Tocris, Catalog # 6640) was used to visualize the endoplasmic reticulum (green) in unfixed PNT2 human prostate epithelial cells. The plasma membrane was stained using CellMask™ (purple).