Analysis Note

ControlHeLa cell lysate

Application

Western Blotting Analysis: A 1:500-2,000 dilution of this antibody detected HSPA5 in 10 µg of NIH/3T3, Jurkat, and A431 cell lysates.

Immunohistochemistry Analysis: A 1:1,000 dilution from a representative lot detected HSPA5 in rat cerebellum, rat pons/midbrain, rat hippocampus, and human thyroid tissues.

Optimal working dilutions must be determined by end user.

Research CategoryApoptosis & Cancer

Anti-HSPA5 Antibody, clone 4E3 is a highly specific mouse monoclonal antibody, that targets GRP-78 & has been tested in western blotting & IHC.

Research Sub CategoryApoptosis - Additional

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

The gene HSPA5 encodes a protein named GRP-78 (78 kDa glucose-regulated protein) also known as BiP (immunoglobulin heavy chain-binding protein) or heat shock 70 kDa protein 5. While originally discovered as one of set of genes that were induced upon cellular starvation and stress, HSPA5 encoded GRP-78 is now known to be an important member of the heat-shock family of proteins. The HSPA5 encoded GRP-78 protein appears to play a role in helping in the assembly of multimeric protein complexes in the endoplasmic reticulum (ER). Cellular localization is in the endoplasmic reticulum lumen and appears as cytoplasmic-like staining. The HSPA5 encoded protein is an essential component of the translocation processes within the endoplasmic reticulum and also functions in the retrograde transport across the ER membrane of incorrectly folded proteins destined to be degraded by the ubiquitin proteasome system. The HSPA5 encoded protein, while widely expressed in all cells types under normal conditions, is particularly induced under conditions of cellular stress, heat, or toxins and conditions which lead to the accumulation of unfolded proteins in the ER. EMD-Millipore's Anti-HSPA5 protein mouse monoclonal antibody, clone 4E3, has been tested by western blot in human HeLa and Jurkat cell lysates as well as mouse NIH3T3 cell lysates. The clone has also been successfully tested in immunohistochemistry in human, paraffin-embedded, thyroid tissues.

Immunogen

Purified recominant fragment of human HSPA5 expressed in E. coli.

Physical form

Mouse monoclonal IgG2b ascitic fluid containing up to 0.1% sodium azide.

Unpurified

Quality

Evaluated by Western Blotting in HeLa cell lysate.

Western Blotting Analysis: A 1:500 dilution of this antibody detected HSPA5 in 10 µg of HeLa cell lysate.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

~72 kDa observed. Uncharacterized bands may appear in some lysate(s).