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Mouse monoclonal antibody to rh c-FOS [2H2]: Affinity purified


Catalogue No. M-1752-100
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:
Batch No. See product label.
Unit size 100 ug
Antigen Full length, E.coli-derived recombinant human c-FOS protein.
Antibody Type Monoclonal
Isotype IgG1
Clone 2H2
Other Names Cellular oncogene fos; G0/G1 switch regulatory protein 7; cFOS
Accession FOS_HUMAN
Produced in Mouse.
Purity Affinity purified.
Applications Immunohistochemistry (IHC) and immunocytochemistry (ICC): 1-2 ug/mL. This antibody has been shown to work on 4% PFA fixed mouse brain sections.

Western blotting (WB): 0.5-1.0 ug/mL. 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 Rat and mouse. Reactivity with other mammalian species not tested, but expected.
Blast URL Click here
Form Lyophilized from a solution containing 3% trehalose in PBS, pH 7.4, without preservatives.
Reconstitution Reconstitute with 100 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)
Mouse monoclonal antibody to rh c-FOS [2H2]: Affinity purified A: Western blot analysis of c-Fos expression in HeLa cells using M-1752-100. HeLa cells were serum-starved for 36 hours (Lane 1). Serum-starved HeLa cells were stimulated with 20% FBS for 2 hours (Lane 2). M-1752-100 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 M-1752-100. c-Fos staining (green) 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 (red) and DAPI (blue).
C: Immunohistochemistry using M-1752-100 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 M-1752-100 (red) and Rabbit polyclonal anti-NeuN/Fox3 (R-3770-100, green) on mouse cortical sections. Neurons positive for c-Fos and Fox3/NeuN appear yellow. The insert shows an enlarged image of staining with M-1752-100. Nuclei were labeled with DAPI (blue).
Mouse monoclonal antibody to rh c-FOS [2H2]: Affinity purified 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.
Western Blotting Method: SDS-PAGE: denaturing and reducing, 4-12% Bis-Tris gel; Transfer: Tris-Glycine (Towbins) 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 4C; Secondary antibodies: anti-rabbit-HRP (1/6000) or anti-mouse-HRP (1/3000), 1 hour at RT; Detection: Chemiluminiscence.
Mouse monoclonal antibody to rh c-FOS [2H2]: Affinity purified Left: Rat hippocampus stained for c-FOS (red) by Immunohistochemistry. Section was co-stained with rabbit antibody to FOX3/NeuN (R-3770-100, green). Blue: DAPI nuclear stain. The hippocampal neurons stain green for FOX3/NeuN and a few also are expressing c-FOS, and thus appear orange. These cells were spontaneously active at the time the animal was sacrificed. Right: Western blot analysis of c-FOS expression (green) in cell lysates. Mouse antibody to c-FOS was used at 1:1,000 dilution. GAPDH (red, lanes 2-5) was used as loading control (rabbit antibody to GAPDH, R-1701-100, 1:20,000). [1] protein standard, [2] HeLa cells in serum free media, [3] HeLa cells stimulated with 20% FBS for 2 hours after 36 hours serum starvation, [4] rat cortical neurons, [5] rat cortical neurons treated with membrane depolarization buffer for 5 hours. Multiple bands at 50-65 kDa in stimulated or treated cell lysates correspond to different forms of the c-Fos protein. The single band at 37 kDa (red) represents GAPDH protein.
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