CST, 5348T, Phospho-c-Fos (Ser32) (D82C12) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying Fos (Ser32) phosphate. Validated for WB,WB,IP,IF,IF,F,ChIP,ChIP. Available in 2 sizes. Highly specific and rigorously validated in-house, Phospho-c-Fos (Ser32) (D82C12) Rabbit Monoclonal Antibody (CST #5348) is ready to ship. Product Usage Information For optimal ChIP and ChIP-seq, 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 Fluorescent Western: 1:1000 Immunoprecipitation: 1:100 Immunofluorescence (Frozen): 1:100 - 1:400 Immunofluorescence (Immunocytochemistry): 1:100 - 1:400 Flow Cytometry (Fixed/Permeabilized): 1:200 - 1:800 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 # 99325 . Protocol Available protocols: Western Blotting, Fluorescent Western, Immunoprecipitation, Immunofluorescence (Frozen), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq Specificity / Sensitivity Phospho-c-Fos (Ser32) (D82C12) Rabbit Monoclonal Antibody detects endogenous levels of c-Fos protein only when phosphorylated on Ser32. This antibody does not cross-react with other Fos proteins, including FosB, FRA1 and FRA2. Non-specific phosphatase-sensitive cytoskeletal staining has been observed by immunofluorescence in P301L Tau Transgenic mouse brain. Species Reactivity: Human, Mouse, Rat Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to Ser32 of human c-Fos protein. Background The Fos family of nuclear oncogenes includes c-Fos, FosB, Fos-related antigen 1 (FRA1), and Fos-related antigen 2 (FRA2) (1). While most Fos proteins exist as a single isoform, the FosB protein exists as two isoforms: full-length FosB and a shorter form, FosB2 (Delta FosB), which lacks the carboxy-terminal 101 amino acids (1-3). The expression of Fos proteins is rapidly and transiently induced by a variety of extracellular stimuli, including growth factors, cytokines, neurotransmitters, polypeptide hormones, and stress. Fos proteins dimerize with Jun proteins (c-Jun, JunB, and JunD) to form Activator Protein-1 (AP-1), a transcription factor that binds to TRE/AP-1 elements and activates transcription. Fos and Jun proteins contain the leucine-zipper motif that mediates dimerization and an adjacent basic domain that binds to DNA. The various Fos/Jun heterodimers differ in their ability to transactivate AP-1 dependent genes. In addition to increased expression, phosphorylation of Fos proteins by Erk kinases in response to extracellular stimuli may further increase transcriptional activity (4-6). Phosphorylation of c-Fos at Ser32 and Thr232 by Erk5 increases protein stability and nuclear localization (5). Phosphorylation of FRA1 at Ser252 and Ser265 by Erk1/2 increases protein stability and leads to overexpression of FRA1 in cancer cells (6). Following growth factor stimulation, expression of FosB and c-Fos in quiescent fibroblasts is immediate, but very short-lived, with protein levels dissipating after several hours (7). FRA1 and FRA2 expression persists longer, and appreciable levels can be detected in asynchronously growing cells (8). Deregulated expression of c-Fos, FosB, or FRA2 can result in neoplastic cellular transformation; however, Delta FosB lacks the ability to transform cells (2,3). Alternate Names activator protein 1; AP-1; C-FOS; cellular oncogene c-fos; Cellular oncogene fos; cfos; FBJ murine osteosarcoma viral (v-fos) oncogene homolog (oncogene FOS); FBJ murine osteosarcoma viral oncogene homolog; FOS; Fos proto-oncogene, AP-1 trancription factor subunit; Fos proto-oncogene, AP-1 transcription factor subunit; G0/G1 switch regulatory protein 7; G0S7; p55; Protein c-Fos; Proto-oncogene c-Fos; v-fos FBJ murine osteosarcoma viral oncogene homolog Specification REACTIVITY: H M R SENSITIVITY: Endogenous MW (kDa): 62 Source/Isotype: Rabbit IgG