CST, 46535T, SMAD4 (D3R4N) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying SMAD4. Validated for WB,IP,IHC,IHC,IF,F,ChIP,ChIP. Available in 2 sizes. Highly specific and rigorously validated in-house, SMAD4 (D3R4N) Rabbit Monoclonal Antibody (CST #46535) is ready to ship. Product Usage Information For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits. Western Blotting: 1:1000 Immunoprecipitation: 1:200 IHC Leica Bond: 1:400 - 1:1600 Immunohistochemistry (Paraffin): 1:200 - 1:800 Immunofluorescence (Immunocytochemistry): 1:400 - 1:1600 Flow Cytometry (Fixed/Permeabilized): 1:400 - 1:1600 Chromatin IP: 1:50 Chromatin IP-seq: 1:50 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. For a carrier free (BSA and azide free) version of this product see product # 73885 . Protocol Available protocols: Western Blotting, Immunoprecipitation, IHC Leica Bond, Immunohistochemistry (Paraffin), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq Specificity / Sensitivity SMAD4 (D3R4N) Rabbit Monoclonal Antibody recognizes endogenous levels of total SMAD4 protein. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with recombinant protein corresponding to human SMAD4 protein. The epitope has been mapped to residues surrounding Ser315. Background Members of the SMAD family of signal transduction molecules are components of a critical intracellular pathway that transmits TGF-β signals from the cell surface into the nucleus. Three distinct classes of SMADs have been defined: the receptor-regulated SMADs (R-SMADs), which include SMAD1, 2, 3, 5, 9; the common-mediator SMAD (co-SMAD), SMAD4; and the antagonistic or inhibitory SMADs (I-SMADs), SMAD6 and 7 (1-5). Activated type I receptors associate with specific R-SMADs and phosphorylate them on a conserved SSXS motif in the carboxy-terminus. Phosphorylated R-SMADS dissociate from the receptor and form a heteromeric complex with SMAD4, initiating translocation of the heteromeric SMAD complex to the nucleus. Once in the nucleus, SMADs recruit a variety of DNA binding proteins that function to regulate transcriptional activity (6-8). Alternate Names deleted in pancreatic carcinoma locus 4; Deletion target in pancreatic carcinoma 4; DPC4; hSMAD4; JIP; MAD homolog 4; MAD, mothers against decapentaplegic homolog 4; MADH4; Mothers against decapentaplegic homolog 4; mothers against decapentaplegic, Drosophila, homolog of, 4; Mothers against DPP homolog 4; MYHRS; SMAD 4; SMAD family member 4; SMAD, mothers against DPP homolog 4; SMAD4 Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 70 Source/Isotype: Rabbit IgG