Application

Immunofluorescence Analysis: A representative lot detected phospho-Smad2 (Ser465, Ser467) in prostate cancer tissue (Yakymovych, I., et. al. (2015). J Cell Biol. 210(2):319-32).

Western Blotting Analysis: A representative lot detected phospho-Smad2 (Ser465, Ser467) in TBP: PC-3U cells stimulated with 5 ng/ml TGF (Yakymovych, I., et. al. (2015). J Cell Biol. 210(2):319-32).

Immunoprecipitation Analysis: A representative lot detected phospho-Smad2 (Ser465, Ser467) in COS cells transfected with constitutively active versions of ALK5 (TLR-I)- and ALK6TL45-mediated phosphorylation (Persson, U., et. al. (1998). FEBS Lett. 434(1-2):83-7).

Anti-phospho-Smad2 (Ser465, Ser467), Cat. No. ABE2872, is a highly specific rabbit polyclonal antibody that targets Mothers against decapentaplegic homolog 2 (Smad2) and has been tested in Immunofluorescence, Immunoprecipitation, and Western Blotting.

Research CategoryEpigenetics & Nuclear Function

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

Mothers against decapentaplegic homolog 2 (UniProt: Q15796; also known as MAD homolog 2, Mothers against DPP homolog 2, JV18-1, Mad-related protein 2, hMAD-2, SMAD family member 2, SMAD 2, Smad2, hSMAD2) is encoded by the SMAD2 (also known as MADH2, MADR2) gene (Gene ID: 4087) in human. Smad2 is a receptor-regulated Smad (R-Smad) that serves as an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor beta (TGF beta) and activin type 1 receptor kinases. It is expressed at high levels in skeletal muscle, endothelial cells, heart, and placenta. It contains one Mad homology (MH) 1 domain (aa 10-176), one MH2 domain (aa 274-467) and a PY motif (aa 221-225). Cytoplasmic Smad2 is directly phosphorylated on Ser465/Ser467 by TGF- receptor kinase I. These C-terminal phosphoserines of R-Smads are recognized by the MH2 domain of Smad4 that enables them to form a heterodimeric complex, which translocates to the nucleus where Smad proteins bind to their cognate DNA binding sites with low affinity. This binding is further enhanced in the presence of transcriptional co-activators. Acetylation Lys-19 by coactivators in response to TGF-beta signaling is shown to increase its transcriptional activity. Upon dephosphorylation by phosphatase PPM1A, Smad2 is released from the Smad2/Smad4 complex and is exported out of the nucleus.

Immunogen

Epitope: C-terminus

KLH-conjugated linear peptide corresponding to 4 amino acids surrounding pSerine from the C-terminus region of human Smad2.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Rabbit polyclonal antiserum with 0.01% sodium azide.

Format: Unpurified

Unpurified

Quality

Evaluated by Western Blotting in HEPG2 stimulated with TGFbeta1 cells.

Western Blotting Analysis: A 1:2,000 dilution of this antibody detected phospho-Smad2 (Ser465, Ser467) in HEPG2 stimulated with TGFbeta1 cells.

Specificity

This rabbit polyclonal antibody detects SMAD2 phosphorylated on Serine 465 and Serine 467.

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

~56 kDa observed; 52.31 kDa calculated. Uncharacterized bands may be observed in some lysate(s).