Anti-SMAD5; clone 3H9

Analysis Note

ControlHeLa, SK-N-SH, PC-12, Jurkat, and K562 cell lysates

Application

Research Sub CategorySignaling Neuroscience

Research CategoryNeuroscience

Detect Smad using this mouse monoclonal antibody, Anti-SMAD5 Antibody, clone 3H9 validated for use in western blotting, Immunofluorescence, IHC & Flow Cytometry.

Immunofluorescence Analysis: A 1:200-1,000 dilution from a representative lot detected SMAD5 in NTERA-2 cells.

Immunohistochemistry Analysis: A 1:200-1,000 dilution from a representative lot detected SMAD5 in human brain and lung cancer tissues.

Flow Cytometry Analysis: A 1:200-400 dilution from a representative lot detected SMAD5 in Jurkat cells.

Optimal working dilutions must be determined by end user.

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

SMAD5, MAD homolog 5, Mothers against DPP homolog 5, JV5-1, SMAD family member 5 encoded by the gene named SMAD5 or MADH5 is a transcriptional cofactor activated by BMP type 1 receptor kinase and like other SMAD family members SMAD5 belongs to the larger TGFbeta superfamily of signaling modulators. SMAD5 forms complexes with SMAD4 when phosphorylated by BMPR1 and migrates to the nucleus. In the absence of phosphorylation SMAD5 is cytoplasmically localized. SMAD5 levels are regulated via ubiquitin mediated proteolysis facilitated by the E3 ligase SMURF1. SMAD5 is involved in many developmental pathways including in hematopoietic cells where SMAD5 activity is involved in the inhibition of the proliferation of hematopoietic progenitor cells. More recent research has shown SMAD5 to be involved in many other pathways too, for instance homozygous mutants die early in development and display a variable phenotype. Moreover SMAD5 is critical for proper granule cell development in the cerebellum. Finally because SMAD5 belongs to the TGFbeta family whose normal function is usually growth suppression, SMAD5 is often mutated in many cancers and its expression is associated with many forms of cancers in a similar fashion to that of its other family members. EMD-Millipore's Anti-SMAD5 mouse monoclonal antibody has been tested in western blot on HeLa, SK-N-SH, PC-12 , Jurkat , and K562 cell lysates and in paraffin embedded immunohistochemistry on human brain and lung cancer tissue sections and in fluorescent immunocytochemistry on NTERA-2 cells in culture and in flow cytometry on Jurkat cells in culture.

Immunogen

Purified recombinant fragment of human SMAD5 expressed in E. Coli.

Physical form

Mouse monoclonal IgG1 ascitic fluid containing up to 0.1% sodium azide.

Unpurified

Quality

Evaluated by Western Blotting in HeLa, SK-N-SH, PC-12, Jurkat, and K562 cell lysates.

Western Blotting Analysis: A 1:500-2,000 dilution of this antibody detected SMAD5 in HeLa, SK-N-SH, PC-12, Jurkat, and K562 cell lysates.

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

~52 kDa observed. Uncharacterized bands may appear in some lysate(s).