Cell Signaling Technology

Product Pathways - MAPK Signaling

Fos Family Antibody Sampler Kit #8333

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb #5348 40 µl W IP IF-IC F ChIP H M R (Hm) (Mk) (B) (Pg) (Hr) 62 Rabbit IgG
FosB (5G4) Rabbit mAb #2251 40 µl W IP IHC-P IF-IC F ChIP H M R 38 FosB2 48 FosB Rabbit IgG
c-Fos (9F6) Rabbit mAb #2250 40 µl W IF-IC F ChIP H M R (Hm) (B) (Pg) 62 Rabbit IgG
FRA1 (D80B4) Rabbit mAb #5281 40 µl W IP H (Mk) 40 Rabbit IgG
Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb #5841 40 µl W ChIP H M R (Mk) (B) (Hr) 40 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  B=Bovine  Pg=Pig  Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the Fos Family Antibody Sampler Kit recognizes endogenous levels of the specific target protein. FosB (5G4) Rabbit mAb detects both FosB and FosB2 isoforms. Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb recognizes endogenous levels of FRA1 protein only when phosphorylated at Ser265. This antibody may also cross-react with phospho-FRA2, but does not cross-react with phospho-c-Fos or phospho-FosB.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and H-4-II-E cells serum-starved overnight and treated with TPA #4174 (4 hrs), using c-Fos (9F6) Rabbit mAb #2250.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells serum-starved overnight and treated with TPA #4174 (4 hrs), or NIH/3T3 cells and C6 cells serum-starved overnight and serum-stimulated (4 hrs), using FosB (5G4) Rabbit mAb #2251.

Western Blotting

Western Blotting

Western blot analysis of extracts from serum-starved HeLa cells, untreated (-) or treated (+) with TPA #4174 (4 hrs), using FRA1 (D80B4) Rabbit mAb #5281 (upper) and β-Actin Antibody #4967 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, serum-starved overnight and then either untreated or treated with TPA #4174 (4 hrs), using Phospho-c-Fos (Ser32) (D82C12) XP® Rabbit mAb #5348 (upper) or c-Fos (9F6) Rabbit mAb #2250 (lower). Antibody phospho-specificity is shown by treating lysates with λ phosphatase.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, serum-starved overnight, and either left untreated or treated with TPA #4174 for 4 hours, using Phospho-FRA1 (Ser265) (D22B1) Rabbit mAb #5841 (upper) and FRA1 (D80B4) Rabbit mAb #5281 (lower). Antibody phospho-specificity is shown by treating lysates with λ phosphatase.

Description

The Fos Family Antibody Sampler Kit provides an economical means to evaluate the Fos family of transcription factors. The kit includes enough antibody to perform four western blot experiments with each primary antibody.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser32 of human c-Fos, Ser265 of human FRA1 protein or with a synthetic peptide derived from human FosB, c-Fos, or FRA1.

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), that 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).

  1. Tulchinsky, E. (2000) Histol. Histopathol. 15, 921-928.
  2. Dobrzanski, P. et al. (1991) Mol. Cell. Biol. 11, 5470-5478.
  3. Nakabeppu, Y. and Nathans, D. (1991) Cell 64, 751-759.
  4. Rosenberger, S.F. et al. (1999) J. Biol. Chem. 274, 1124-1130.
  5. Sasaki, T. et al. (2006) Mol. Cell 24, 63-75.
  6. Basbous, J. et al. (2007) Mol. Cell. Biol. 27, 3936-3950.
  7. Kovary, K. and Bravo, R. (1991) Mol. Cell. Biol. 11, 2451-2459.
  8. Kovary, K. and Bravo, R. (1992) Mol. Cell. Biol. 12, 5015-5023.

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