Skip to main content

Rabbit Recombinant Monoclonal A RAF antibody. Suitable for Flow Cyt (Intra), WB, ICC/IF and reacts with Mouse, Human, Rat, Transfected cell lysate - Mouse samples.

Be the first to review this product! Submit a review

Images

Immunocytochemistry/ Immunofluorescence - Anti-A-Raf antibody [EPR25405-17] (AB314539), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-A-Raf antibody [EPR25405-17] (AB314539), expandable thumbnail
  • Western blot - Anti-A-Raf antibody [EPR25405-17] (AB314539), expandable thumbnail
  • Western blot - Anti-A-Raf antibody [EPR25405-17] (AB314539), expandable thumbnail
  • Western blot - Anti-A-Raf antibody [EPR25405-17] (AB314539), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
Flow Cyt (Intra)WBICC/IFIHC-PIP
Human
Expected
Tested
Expected
Not recommended
Not recommended
Mouse
Tested
Tested
Tested
Not recommended
Not recommended
Rat
Expected
Tested
Expected
Not recommended
Not recommended
Transfected cell lysate - Mouse
Not recommended
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Mouse
Dilution info
1/50
Notes

-

Expected
Expected

Species
Human, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Transfected cell lysate - Mouse
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Rat
Dilution info
1/1000
Notes

-

Species
Transfected cell lysate - Mouse
Dilution info
1/1000
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/100
Notes

-

Expected
Expected

Species
Human, Rat
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Transfected cell lysate - Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat, Transfected cell lysate - Mouse
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Human, Rat, Transfected cell lysate - Mouse
Dilution info
-
Notes

-

Associated Products

Select an associated product type

1 product for Alternative Version

Target data

Function

Involved in the transduction of mitogenic signals from the cell membrane to the nucleus. May also regulate the TOR signaling cascade (By similarity). Phosphorylates PFKFB2 (By similarity).

Alternative names

A-raf, Araf1, Araf, Serine/threonine-protein kinase A-Raf, Proto-oncogene A-Raf

Recommended products

Rabbit Recombinant Monoclonal A RAF antibody. Suitable for Flow Cyt (Intra), WB, ICC/IF and reacts with Mouse, Human, Rat, Transfected cell lysate - Mouse samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR25405-17
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The A-Raf protein also called ARAF is a serine/threonine-protein kinase with a molecular mass of about 68 kDa. Scientists frequently refer to it with names like A-Raf or anti A-Raf when discussing its function. A-Raf is expressed in various tissues with higher levels found in the urogenital system and the central nervous system. This protein plays an important role in cellular signal transduction processes influencing cell proliferation and differentiation through phosphorylation activity.

Biological function summary

The A-Raf protein participates in the regulation of cell growth and survival. It forms part of the MAPK/ERK signaling cascade which is essential for transmitting mitogenic signals from the cell surface to the nucleus. A-Raf acts as a modulator within this complex impacting the overall signaling output. The protein is intimately involved in the regulation of the developmental processes influencing cell fate and function.

Pathways

The A-Raf protein is central to the MAPK/ERK pathway a critical signaling pathway involved in the regulation of cellular responses to growth signals. This pathway includes key components like the proteins B-Raf and C-Raf with A-Raf playing specific roles in the modulation and fine-tuning of this signal transmission. It helps mediate signals that control cell division differentiation and secretion ensuring appropriate cellular responses to external stimuli.

Associated diseases and disorders

Aberrations in A-Raf functions have been linked to cancers particularly urogenital cancers due to its role in uncontrolled cell proliferation. Faulty A-Raf activity can also contribute to neurological disorders because of its expression in the central nervous system and involvement in neuronal signaling pathways. In these contexts it can interact with proteins like B-Raf which are important for maintaining normal cellular signaling and preventing tumorigenesis and neurodegenerative disorders.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-A-Raf antibody [EPR25405-17] (ab314539), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-A-Raf antibody [EPR25405-17] (ab314539)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling A-Raf with ab314539 at 1/100 (5.35 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1007 (2ug/ml) dilution (Green). Confocal image showing mitochondrial staining in NIH/3T3 cells. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1007 (2ug/ml) dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-A-Raf antibody [EPR25405-17] (ab314539), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-A-Raf antibody [EPR25405-17] (ab314539)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MEF (mouse embryo fibroblast) cells labelling A-Raf with ab314539 at 1/100 (5.35 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1006 (2ug/ml) dilution (Green). Confocal image showing mitochondrial staining in MEF cells. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1006 (2ug/ml) dilution.

  • Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539), expandable thumbnail

    Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    This antibody does not cross-react with mouse B-Raf and c-Raf.
    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
    In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

    All lanes: Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539) at 1/1000 dilution

    Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 5 µg

    Lane 2: 293T cells transfected with a mouse A-Raf expression vector containing a His-tag, whole cell lysate at 5 µg

    Lane 3: 293T cells transfected with a mouse c-Raf expression vector containing a His-tag, whole cell lysate at 5 µg

    Lane 4: 293T cells transfected with a mouse B-Raf expression vector containing a His-tag, whole cell lysate at 5 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Observed band size: 68 kDa

    Exposure time: 3s

  • Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539), expandable thumbnail

    Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    The identity of the bands between 10 kDa and 68 kDa are unknown.
    Exposure time: Lane 1: 26 seconds;
    Lanes 2-3: 180 seconds.

    All lanes: Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539) at 1/1000 dilution

    Lane 1: Mouse epididymis tissue lysate at 20 µg

    Lane 2: Mouse prostate tissue lysate at 20 µg

    Lane 3: C2C12 (mouse myoblast) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 68 kDa

  • Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539), expandable thumbnail

    Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

    All lanes: Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539) at 1/1000 dilution

    Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Lane 3: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

    Lane 4: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 68 kDa

    Exposure time: 92s

  • Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539), expandable thumbnail

    Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST
    The identity of the lower MW band at approximately 42 kDa (in lane 3) is unknown.
    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    All lanes: Western blot - Anti-A-Raf antibody [EPR25405-17] (ab314539) at 1/1000 dilution

    Lane 1: NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

    Lane 2: NIH/3T3 transfected with siRNA specifically targeti A-Raf whole cell lysate at 20 µg

    Lane 3: MEF (mouse embryo fibroblast) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 68 kDa

    Exposure time: 180s

  • Flow Cytometry (Intracellular) - Anti-A-Raf antibody [EPR25405-17] (ab314539), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-A-Raf antibody [EPR25405-17] (ab314539)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MEF (mouse embryo fibroblast) cells labelling A-Raf with ab314539 at 1/50 dilution (1 ug, Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

  • Flow Cytometry (Intracellular) - Anti-A-Raf antibody [EPR25405-17] (ab314539), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-A-Raf antibody [EPR25405-17] (ab314539)

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling A-Raf with ab314539 at 1/50 dilution (1 ug, Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com