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Rabbit polyclonal antibody to rh c-FOS: Affinity purified

$297.00USD


Catalogue No. R-1751-50
Description FUNCTION: Nuclear phosphoprotein which forms a tight but non-covalently linked complex with the JUN/AP-1 transcription factor. In the heterodimer, FOS and JUN/AP-1 basic regions each seems to interact with symmetrical DNA half sites. On TGF-beta activation, forms a multimeric SMAD3/SMAD4/JUN/FOS complex at the AP1/SMAD-binding site to regulate TGF-beta-mediated signaling. Has a critical function in regulating the development of cells destined to form and maintain the skeleton. It is thought to have an important role in signal transduction, cell proliferation and differentiation. In growing cells, activates phospholipid synthesis, possibly by activating CDS1 and PI4K2A. This activity requires Tyr-dephosphorylation and association with the endoplasmic reticulum. SUBUNIT: Heterodimer. Interacts with DSIPI; this interaction inhibits the binding of active AP1 to its target DNA. Interacts with MAFB. SUBCELLULAR LOCATION: Nucleus. INDUCTION: C-fos expression increases upon a variety of stimuli, including growth factors, cytokines, neurotransmitters, polypeptide hormones, stress and cell injury. SIMILARITY: Belongs to the bZIP family. Fos subfamily. SIMILARITY: Contains 1 bZIP domain (Ref: uniprot.org).
Batch No. See product label.
Unit size 50 ug
Antigen Full length, E.coli-derived recombinant human c-FOS protein.
Antibody Type Polyclonal.
Other Names Cellular oncogene fos; G0/G1 switch regulatory protein 7; cFOS
Accession FOS_HUMAN
Produced in Rabbit.
Purity Affinity purified.
Applications Immunocytochemistry (ICC): 1:2,000-1:10,000.

Immunohistochemistry (IHC): 1:20,000-1:50,000. This antibody has been shown to work on 4% PFA fixed mouse brain sections. Note that non-specific staining has been observed on tissue sections when using this antibody at dilutions of 1:5,000 or lower. Click here for instructions on use of this antibody in IHC on free-floating brain sections.

Western blotting (WB): 1:1,000-1:2,000. This antibody detects bands between 50-65 kDa, which only appear in stimulated cells.

Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
Specificity Human.
Cross-reactivity Horse, cow, pig, chicken, rat, mouse.
Blast URL Click here
Form Lyophilized from a solution containing 3% trehalose in PBS, pH 7.4, without preservatives.
Reconstitution Reconstitute with 50 uL sterile-filtered, ultrapure water, to achieve an antibody concentration of 1 mg/mL. Centrifuge briefly to remove any insoluble material.
Storage Store lyophilised, unopened vial at 2-8C or lower. After reconstitution, prepare aliquots and store at -20C for a higher stability. Avoid freeze-thaw cycles.
Expiry Date 12 months after purchase if unopened and stored as indicated.
General References Vanstraaten et al (1983) Proc. Natl. Acad. Sci. 80: 3183 (Original molecular c-fos sequence paper)

Minson J. et al. (1994) Brain Res. 646: 44-52 (Early IHC localization paper)
Images (click to zoom)
Rabbit polyclonal antibody to rh c-FOS: Affinity purified Analysis of c-Fos expression in cultured HeLa cells by Immunocytochemistry (A-B), and in mouse hippocampus (C) or olfactory bulb sections (D) by Immunohistochemistry. HeLa cells were kept in serum-free medium for 36 hours, before cells were stimulated with 20% FBS for 30 minutes (B), while control cells (A) were treated with PBS as a control. The rabbit anti-c-Fos antibody (red, 1:5,000) labels nuclei of stimulated cells, while DAPI (blue) stains all cell nuclei independent of their activity stage. Green: mouse anti-tubulin antibody. Mouse hippocampus (C) and olfactory bulb sections (D) were stained with rabbit anti-c-Fos antibody (red, 1:20,000) and mouse antibody to NF-L (green). The c-FOS antibody stains only nuclei of spontaneously active neurons. NF-L is expressed in axons of neuronal cells. Blue: Cell nuclei stained with DAPI. IHC Method: Following transcardial perfusion of mouse with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45 uM, and free-floating sections were stained with above antibodies.
Rabbit polyclonal antibody to rh c-FOS: Affinity purified A: Western blot analysis of c-Fos expression in HeLa cells using R-1751-50. HeLa cells were serum-starved for 36 hours (Lane 1). Serum-starved HeLa cells were stimulated with 20% FBS for 2 hours (Lane 2). R-1751-50 recognizes bands in the range of 50-65 kDa, which represent multiple forms of c-Fos. A loading control was performed by stripping and re-probing the membrane with a monoclonal antibody against GAPDH, M-1376-250.
B: Immunofluorescence staining of HeLa cells with R-1751-50. c-Fos staining (red) only localizes in the nuclei of 20% FBS stimulated cells (bottom panel), but not in un-stimulated cells (top panel). Cells were counter-stained with Chicken polyclonal antibody against vimentin, C-1409-50 (green) and DAPI (blue).
C: Immunohistochemistry using R-1751-50 on 4% PFA transcardial-perfused mouse brain sections (45 uM thickness). c-FOS immunoreactive cells (dark colour, localized in cell nucleus) were visualized using a standard HRP-DAB (horseradish peroxidase-3,3\'-diaminobenzidine) staining technique. D: Immunohistochemistry using R-1751-50 (red) and Mouse monoclonal anti-NeuN/Fox3 (M-377-100, green) on mouse cortical sections. Neurons positive for c-Fos and Fox3/NeuN appear yellow. The insert shows an enlarged image of staining with R-1751-50. Nuclei were labeled with DAPI (blue).
Rabbit polyclonal antibody to rh c-FOS: Affinity purified A: Western blot analysis of c-Fos expression in rat C6 cells (40 ug lysate loading per lane). C6 cells were serum-starved for 20 hours and then stimulated with 20% FBS for 2 hrs (+). Control cells were left in serum-depleted culture medium (-). M-1752-100 and R-1751-50 detect a strong band at 50 kDa in stimulated cells but not in control cells, representing increased c-FOS expression.
B:Western blot analysis of c-Fos expression in PC12 cell lysates (Lane 1, 20 μg), mouse brain (Lane 2, 50 μg) and rat spinal cord homogenate (Lane 3, 50 μg). R-1751-50 detects multiple c-Fos isoforms between ~50-65 kDa which are due to post-translational modifications. In mouse brain, R-1751-50 detects a common c-Fos degradation product (~41 kDa) which appears to be lysate and preparation dependent.
Western Blotting Method: SDS-PAGE: denaturing and reducing, 4-12% Bis-Tris gel; Transfer: Tris-Glycine (Towbin’s) buffer with 20% methanol; Membrane: PVDF (0.45 μm); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibodies: R-1751-50 (1 μg/mL), M-1752-100 (1 μg/mL), incubation overnight at 4°C; Secondary antibodies: anti-rabbit-HRP (1/6000) or anti-mouse-HRP (1/3000), 1 hour at RT; Detection: Chemiluminiscence.
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