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Rabbit Recombinant Monoclonal RAF1 phospho S259 antibody. Suitable for Dot, WB and reacts with Synthetic peptide, Mouse, Rat, Human samples. Cited in 17 publications.

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Images

Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539), expandable thumbnail
  • Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539), expandable thumbnail
  • Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539), expandable thumbnail
  • Dot Blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539), expandable thumbnail
  • Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

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
IPFlow CytDotWBICC/IF
Human
Not recommended
Not recommended
Expected
Tested
Not recommended
Mouse
Not recommended
Not recommended
Expected
Tested
Not recommended
Rat
Not recommended
Not recommended
Expected
Tested
Not recommended
Synthetic peptide
Not recommended
Not recommended
Tested
Not recommended
Not recommended

Not recommended
Not recommended

Species
Mouse, Rat, Human, Synthetic peptide
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Human, Synthetic peptide
Dilution info
-
Notes

-

Tested
Tested

Species
Synthetic peptide
Dilution info
-
Notes

-

Expected
Expected

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

-

Tested
Tested

Species
Mouse
Dilution info
1/1000 - 1/5000
Notes

-

Species
Rat
Dilution info
1/1000 - 1/5000
Notes

-

Species
Human
Dilution info
1/1000 - 1/5000
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat, Synthetic peptide
Dilution info
-
Notes

-

Associated Products

Select an associated product type

13 products for Alternative Product

Target data

Function

Serine/threonine-protein kinase that acts as a regulatory link between the membrane-associated Ras GTPases and the MAPK/ERK cascade, and this critical regulatory link functions as a switch determining cell fate decisions including proliferation, differentiation, apoptosis, survival and oncogenic transformation. RAF1 activation initiates a mitogen-activated protein kinase (MAPK) cascade that comprises a sequential phosphorylation of the dual-specific MAPK kinases (MAP2K1/MEK1 and MAP2K2/MEK2) and the extracellular signal-regulated kinases (MAPK3/ERK1 and MAPK1/ERK2). The phosphorylated form of RAF1 (on residues Ser-338 and Ser-339, by PAK1) phosphorylates BAD/Bcl2-antagonist of cell death at 'Ser-75'. Phosphorylates adenylyl cyclases: ADCY2, ADCY5 and ADCY6, resulting in their activation. Phosphorylates PPP1R12A resulting in inhibition of the phosphatase activity. Phosphorylates TNNT2/cardiac muscle troponin T. Can promote NF-kB activation and inhibit signal transducers involved in motility (ROCK2), apoptosis (MAP3K5/ASK1 and STK3/MST2), proliferation and angiogenesis (RB1). Can protect cells from apoptosis also by translocating to the mitochondria where it binds BCL2 and displaces BAD/Bcl2-antagonist of cell death. Regulates Rho signaling and migration, and is required for normal wound healing. Plays a role in the oncogenic transformation of epithelial cells via repression of the TJ protein, occludin (OCLN) by inducing the up-regulation of a transcriptional repressor SNAI2/SLUG, which induces down-regulation of OCLN. Restricts caspase activation in response to selected stimuli, notably Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation.

Alternative names

RAF, RAF1, RAF proto-oncogene serine/threonine-protein kinase, Proto-oncogene c-RAF, Raf-1, cRaf

Recommended products

Rabbit Recombinant Monoclonal RAF1 phospho S259 antibody. Suitable for Dot, WB and reacts with Synthetic peptide, Mouse, Rat, Human samples. Cited in 17 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR3433(2)
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

Raf1 also known as c-Raf or Raf-1 is a serine/threonine-protein kinase that plays an important role in cell division differentiation and survival. Its molecular weight is approximately 74 kDa. Raf1 is part of the MAPK/ERK signaling pathway and is ubiquitously expressed in many tissues including the brain heart and liver. As a component of the signaling cascade its activation translates extracellular signals into intracellular responses.

Biological function summary

Raf1 regulates important cellular processes by activating downstream kinases in response to external stimuli. Raf1 forms a complex with other proteins such as Ras facilitating its role as an essential component of the MAPK/ERK pathway. Upon activation by Ras Raf1 phosphorylates and activates MEK1 and MEK2 which in turn activate the ERK1 and ERK2. This signaling axis is involved in controlling gene expression and cellular proliferation.

Pathways

Raf1 is integrally involved in the MAPK/ERK pathway which is critical for transducing signals from growth factors and mitogens. It relates closely with proteins such as Ras MEK and ERK in this pathway. The pathway is important for regulating cellular responses to various stimuli and is particularly involved in processes such as cell cycle control and apoptosis.

Associated diseases and disorders

Raf1 has associations with certain types of cancer and Noonan syndrome. In various cancers mutations or dysregulation of Raf1 or related components such as Ras can lead to uncontrolled cellular proliferation. In Noonan syndrome Raf1 mutations result in anomalies in the Ras/MAPK pathway which can impact normal development. Raf1’s interactions in these pathways indicate its relevance as a potential target for therapeutic intervention.

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.

6 product images

  • Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539), expandable thumbnail

    Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539)

    All lanes: Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution

    Lane 1: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

    Lane 2: NIH/3T3 (Mouse embryonic fibroblast) treated with 5uM forskolin for 10 minutes whole cell lysates at 15 µg

    Lane 3: NIH/3T3 (Mouse embryonic fibroblast) treated with 5uM forskolin for 10 minutes whole cell lysates, then incubated with phosphatase at 15 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

    Predicted band size: 73 kDa

    Observed band size: 73 kDa

  • Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539), expandable thumbnail

    Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539)

    All lanes: Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution

    Lane 1: C6 (Rat glial tumor glial cell) whole cell lysates at 15 µg

    Lane 2: C6 (Rat glial tumor glial cell) treated with 30uM forskolin for 20 minutes whole cell lysates at 15 µg

    Lane 3: C6 (Rat glial tumor glial cell) treated with 30uM forskolin for 20 minutes whole cell lysates, then incubated with phosphatase at 15 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

    Predicted band size: 73 kDa

    Observed band size: 73 kDa

  • Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539), expandable thumbnail

    Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/5000 dilution

    Lane 1: HEK293 whole cell lysate at 15 µg

    Lane 2: HEK293 whole cell lysate treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour at 15 µg

    Lane 3: HEK293 whole cell lysate treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour then incubated with phosphatase at 15 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

    Predicted band size: 73 kDa

    Observed band size: 73 kDa

    Exposure time: 1s

  • Dot Blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539), expandable thumbnail

    Dot Blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539)

    Dot blot analysis of Raf1 (pS259) peptide (Lane 1) and Raf1 non-phospho peptide (Lane 2) labelling Raf1 (pS259) with ab173539 at a dilution of 1/1000. Goat Anti-Rabbit IgG H&L (HRP) ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539), expandable thumbnail

    Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539)

    All lanes: Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) at 1/1000 dilution

    Lane 1: 293T cell lysates at 10 µg

    Lane 2: 293T cell lysates treated with Okadaic Acid + Calyculin A at 10 µg

    Predicted band size: 73 kDa

  • Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539), expandable thumbnail

    Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539)

    All lanes: Purified at 1/5000 dilution

    Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 15 µg

    Lane 2: HEK-293 (Human embryonic kidney epithelial cell) treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour whole cell lysates at 15 µg

    Lane 3: HEK-293 (Human embryonic kidney epithelial cell) treated with 100nM Phorbol-12-myristate-13-acetate for 1 hour whole cell lysates, then incubated with phosphatase at 15 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

    Predicted band size: 73 kDa

    Observed band size: 73 kDa

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Product protocols

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

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