Anti-Cellular oncogene fos (c-Fos) Antibody (2H2)

Name: Anti-Cellular oncogene fos (c-Fos) Antibody
Brand: Biosensis
SKU: M-1752-100
Price: 327.0 USD
Availability: InStock

Our Anti-Cellular oncogene fos (c-Fos) mouse monoclonal primary antibody detects bovine, chicken, horse, human, mouse, pig, and rat Cellular oncogene fos (c-Fos), and is affinity-purified. It is validated for use in ICC, IHC-Frozen.

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).

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 (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.

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.

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).

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Species Reactivity

Bovine, Chicken, Horse, Human, Mouse, Pig, Rat

Applications

ICC, IHC

Host Species

Mouse

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Data Sheet

SKU: M-1752-100

Volume/Size: 100 µg

Variant

Price:

$327.00

Quantity:

Product Details

Target

Cellular oncogene fos (c-Fos)

Scientific Background

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).

Format

Affinity-purified

Clonality

Monoclonal

Clone

2H2

Isotype

IgG1

Applications

ICC, IHC

Host Species

Mouse

Molecular Weight

see application details

Immunogen

Full length, E.coli-derived recombinant human c-FOS protein.

Immunogen Species

Human

Species Reactivity

Bovine, Chicken, Horse, Human, Mouse, Pig, Rat

Storage

Spin vial briefly before opening. Reconstitute with 100 µL sterile-filtered, ultrapure water to achieve a 1 mg/mL concentration. Centrifuge to remove any insoluble material. Store lyophilized, unopened vial at 2-8°C or lower. After reconstitution, prepare aliquots and store at -20°C for a higher stability. Avoid freeze-thaw cycles.

Physical State

Lyophilized

Production Notes

Protein G purified

Buffer

Lyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, and sodium azide

Dilution Ranges

WB: 0.5-1 µg/mL
IHC: 1-2 µg/mL
ICC: 1-2 µg/mL

Application Details

IHC: This antibody has been shown to work on 4% PFA fixed mouse brain sections.

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

Conjugate

Unconjugated

Specificity

Human. Horse, cow, pig, chicken, rat, mouse.

Usage Statement

For research use only.

Country of Origin

United States

Expiration

12 months after date of receipt (unopened vial).

Synonyms

Cellular oncogene fos; G0/G1 switch regulatory protein 7; cFOS

UniProt ID

P01100

Shipping

25°C (ambient)

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Anti-Cellular oncogene fos (c-Fos) Antibody (2H2)

SKU: M-1752-100

Product Details

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