AB173539

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

RabMAb
Recombinant
Lab Essentials
What is this?

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(22 Publications)

Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) is a rabbit monoclonal antibody detecting Raf1 in Western Blot, Dot Blot. Suitable for Human, Mouse, Rat.

- Biophysical QC for unrivalled batch-batch consistency
- Over 10 publications

View Alternative Names

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

6 Images

Lab

Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539)

ab181602 was used as GAPDH loading control. ab181115 was used to detect Raf1. ab150365 was used to detect Raf1 phospho S43.

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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 73 kDa

false

Lab

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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 73 kDa

false

Exposure time: 1s

Lab

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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 73 kDa

false

Lab

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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 73 kDa

false

Supplier Data

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

false

Lab

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. 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.

Carrier free

Anti-Raf1 (phospho S259) antibody [EPR3433(2)] - BSA and Azide free

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR3433(2)

Isotype

IgG

Carrier free

Reacts with

Mouse, Rat, Human

Applications

Dot, WB

applications

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Reactivity data

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Product details

What is this antibody validated in?
Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (ab173539) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Dot Blot in Human, Mouse, Rat samples.

What is the molecular weight of Raf1?
Anti-Raf1 (phospho S259) [EPR3433(2)] (ab173539) specifically detects a band for Raf1 (UniProt: P04049) at a molecular weight of 73kDa.

Trusted by the scientific community
Anti-Raf1 (phospho S259) [EPR3433(2)] (ab173539) was first used in a scientific publication in 2013 and has been cited over 10 times in peer-reviewed journals.

Trial sizes available!
Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products
We have a range of other formats of antibody clone [EPR3433(2)] also available for your convenience: ab173539, Carrier free - ab240957

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

What are the advantages of a recombinant monoclonal antibody?
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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

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

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Supplementary information

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

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.

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.

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Product protocols

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

Visit the General protocols
Visit the Troubleshooting

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Target data

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.

See full target information RAF1 phospho S259

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Publications (22)

Recent publications for all applications. Explore the full list and refine your search

Cancer innovation 3:e117 PubMed38947754

2024

MAPK4 facilitates angiogenesis by inhibiting the ERK pathway in non-small cell lung cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Jing Chen,Jing Yang,Yufang Liu,Xu Zhao,Juanjuan Zhao,Lin Tang,Mengmeng Guo,Ya Zhou,Chao Chen,Dongmei Li,Zhenke Wen,Guiyou Liang,Lin Xu

Journal of traditional Chinese medicine = Chung i tsa chih ying wen pan 44:448-457 PubMed38767628

2024

Optimized new Shengmai powder inhibits myocardial fibrosis in heart failure by regulating the rat sarcoma/rapidly accelerated fibrosarcoma/mitogen-activated protein kinase kinase/extracellular regulated protein kinases signaling pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Zhang Zeyu,Jia Zhuangzhuang,Song Yuwei,Zhang Xuan,Wang Ci,Wang Shuai,Zhang Peipei,Ren Qiuan,Wang Xianliang,Mao Jingyuan

Life science alliance 7: PubMed38499327

2024

SIRT4 as a novel interactor and candidate suppressor of C-RAF kinase in MAPK signaling.

Applications

Unspecified application

Species

Unspecified reactive species

Mehrnaz Mehrabipour,Saeideh Nakhaei-Rad,Radovan Dvorsky,Alexander Lang,Patrick Verhülsdonk,Mohammad R Ahmadian,Roland P Piekorz

Journal of traditional and complementary medicine 13:600-610 PubMed38020551

2023

Xiaozheng pill exerts an anti-mammary hyperplasia effect through Raf/ERK/ELK and HIF-1α/bFGF pathways.

Applications

Unspecified application

Species

Unspecified reactive species

Yu-Fei Liu,Tian An,Hong Yu,Ying-Yi Fan,Xiao-Hua Pei

Journal of experimental & clinical cancer research : CR 42:45 PubMed36759880

2023

Procoxacin bidirectionally inhibits osteoblastic and osteoclastic activity in bone and suppresses bone metastasis of prostate cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Depei Kong,Chen Ye,Chenxi Zhang,Xiaochen Sun,Fubo Wang,Rui Chen,Guangan Xiao,Shipeng He,Jianrong Xu,Xiwu Rao,Jianzhong Ai,Xu Gao,Hong Li,Li Su

Nature structural & molecular biology 29:966-977 PubMed36175670

2022

Structure of the SHOC2-MRAS-PP1C complex provides insights into RAF activation and Noonan syndrome.

Applications

Unspecified application

Species

Unspecified reactive species

Daniel A Bonsor,Patrick Alexander,Kelly Snead,Nicole Hartig,Matthew Drew,Simon Messing,Lorenzo I Finci,Dwight V Nissley,Frank McCormick,Dominic Esposito,Pablo Rodriguez-Viciana,Andrew G Stephen,Dhirendra K Simanshu

Oncology letters 24:223 PubMed35720479

2022

BPTF activates the MAPK pathway through coexpression with Raf1 to promote proliferation of T-cell lymphoma.

Applications

Unspecified application

Species

Unspecified reactive species

Dongyu Bai,Yong Zhou,Fayan Shen,Dehong Gao,Wenhao Suo,Haiping Zhang,Heng Li

Journal of Cancer 13:1972-1984 PubMed35399719

2022

METTL3 Facilitates Tumor Progression by COL12A1/MAPK Signaling Pathway in Esophageal Squamous Cell Carcinoma.

Applications

Unspecified application

Species

Unspecified reactive species

Jiali Li,Zhenhua Li,Yanzhao Xu,Chao Huang,Baoen Shan

Protein expression and purification 193:106061 PubMed35131438

2022

Polycistronic baculovirus expression of SUGT1 enables high-yield production of recombinant leucine-rich repeat proteins and protein complexes.

Applications

Unspecified application

Species

Unspecified reactive species

Kelly Snead,Vanessa Wall,Hannah Ambrose,Dominic Esposito,Matthew Drew

Cancer cell international 22:32 PubMed35042538

2022

4,5-Dimethoxycanthin-6-one is a novel LSD1 inhibitor that inhibits proliferation of glioblastoma cells and induces apoptosis and pyroptosis.

Applications

Unspecified application

Species

Unspecified reactive species

Wei Li,Bai-Sheng Huang,Yuan-Yuan Xiong,Li-Jian Yang,Li-Xiang Wu

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

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

Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539)

Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539)

Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539)

Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539)

Western blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539)

Dot Blot - Anti-Raf1 (phospho S259) antibody [EPR3433(2)] (AB173539)

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Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

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Secondary Antibodies

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Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (22)

Product promise