Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb (#5348) Datasheet with Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:

WB, IP, IF-F, IF-IC, FC-FP, ChIP, ChIP-seq

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

Source/Isotype:

Rabbit IgG

UniProt ID:

#P01100

Entrez-Gene Id:

2353

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 10⁶ cells) per IP. This antibody has been validated using SimpleChIP^® Enzymatic Chromatin IP Kits.

Application	Dilution
Western Blotting	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.

Specificity / Sensitivity

Phospho-c-Fos (Ser32) (D82C12) XP^® Rabbit mAb detects endogenous levels of c-Fos protein only when phosphorylated on Ser32. The 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

Species predicted to react based on 100% sequence homology

Hamster, Monkey, Bovine, Pig, Horse

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).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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