ARAF Recombinant Monoclonal Antibody

Datasheet

Code	CSB-RA044921A0HU
Size	US$210
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Image	IHC image of CSB-RA044921A0HU diluted at 1:100 and staining in paraffin-embedded human cervical cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. IHC image of CSB-RA044921A0HU diluted at 1:100 and staining in paraffin-embedded human thyroid tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. Overlay Peak curve showing Hela cells stained with CSB-RA044921A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/110⁶cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/110⁶cells) used under the same conditions. Acquisition of >10,000 events was performed.
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Target Background

Product Details

Uniprot No.

P10398

Target Names

ARAF

Alternative Names

Serine/threonine-protein kinase A-Raf (EC 2.7.11.1) (Proto-oncogene A-Raf) (Proto-oncogene A-Raf-1) (Proto-oncogene Pks), ARAF, ARAF1 PKS PKS2

Species Reactivity

Human

Immunogen

A synthesized peptide from human ARAF protein

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

18D4

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, IHC, FC

Recommended Dilution

Application	Recommended Dilution
IHC	1:50-1:200
FC	1:50-1:200

Protocols

ELISA Protocol
Immunohistochemistry (IHC) Protocol
Flow Cytometry (FC) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Description

ARAF (A-Raf proto-oncogene, serine/threonine kinase) functions as a critical upstream activator in the MAPK/ERK signaling cascade, playing essential roles in cell proliferation, differentiation, and survival. As one of three RAF family kinases, ARAF contributes to signal transduction from cell surface receptors to the nucleus, making it a significant target for researchers investigating oncogenic signaling pathways, developmental biology, and therapeutic resistance mechanisms in cancer.

This recombinant monoclonal antibody, clone 18D4, offers the reproducibility and consistency that demanding research applications require. Generated against a synthetic peptide derived from human ARAF protein, the antibody is produced using recombinant technology, ensuring sequence-defined specificity and eliminating the lot-to-lot variability often encountered with traditional hybridoma-derived antibodies. This consistency is particularly valuable for longitudinal studies or multi-site collaborations where experimental reproducibility is paramount.

Validation testing demonstrates reliable performance across multiple platforms. Immunohistochemistry studies confirm clear detection in paraffin-embedded human cervical cancer and thyroid tissue sections at 1:100 dilution using citrate buffer antigen retrieval, providing researchers flexibility when examining ARAF expression patterns in clinical specimens. Flow cytometry analysis using HeLa cells shows distinct positive signal separation from isotype control, enabling quantitative assessment of ARAF levels in cell populations at dilutions between 1:50 and 1:200.

The unconjugated format and affinity-purified rabbit IgG preparation allow integration into existing detection workflows using standard secondary antibody systems. Supplied in a glycerol-containing buffer for optimal stability, this antibody serves researchers exploring RAF-MEK-ERK pathway dynamics, tumor progression mechanisms, and targeted therapy development.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Customer Reviews and Q&A

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Target Background

Function

Involved in the transduction of mitogenic signals from the cell membrane to the nucleus. May also regulate the TOR signaling cascade.; Serves as a positive regulator of myogenic differentiation by inducing cell cycle arrest, the expression of myogenin and other muscle-specific proteins, and myotube formation.

Gene References into Functions

The constitutive or induced re-localization of A-Raf to the plasma membrane compromises its ability to efficiently sequester and inactivate MST2, thus rendering cells susceptible to apoptosis PMID: 26891695
This review discusses the regulation of A-Raf protein expression, and the roles of A-Raf in apoptosis and cancer, with a special focus on its role in resistance to Raf inhibitors. [review] PMID: 26508523
analysis of FGFR2-PPHLN1 fusion and ARAF mutations in intrahepatic cholangiocarcinoma PMID: 25608663
Aberrant expression of A-, B-, and C-RAF, and COT is frequent in PTC; increased expression of COT is correlated with recurrence of PTC. PMID: 25674762
Dimerization of the kinase ARAF promotes MAPK pathway activation and cell migration. PMID: 25097033
The study identifies somatic activating ARAF mutations in Langerhans cell histiocytosis. PMID: 24652991
Galpha12-ARAF-ERK pathway stimulates RFFL transcription through the transcription factor c-Myc. PMID: 24114843
show that Araf antagonizes mesendoderm induction and patterning activity of Nodal/Smad2 signals in vertebrate embryos by directly inhibiting Smad2 signalling PMID: 23591895
study investigated role of ARAF in cancer cell signaling and examined the role of ARAF in mediating paradoxical activation of the MAPK pathway in cells treated with RAF inhibitors; ARAF seems to stabilize BRAF:CRAF complexes in cells treated with RAF inhibitors and regulate cell signaling in a subtle manner to ensure signaling efficiency PMID: 22926515
Ras pathway activation via EGF treatment induced strong binding between B-Raf and C-Raf and a low level of binding between B-Raf and A-Raf. PMID: 23352452
hnRNP H blocks MST2-mediated apoptosis in cancer cells by regulating A-Raf transcription. PMID: 20145135
In a two-hybrid screen of human fetal liver cDNA library, TH1 was detected as a new interaction partner of A-Raf; this specific interaction may have played a critical role in the activation of A-Raf. PMID: 11952167
A-Raf kinase is negatively regulated by trihydrophobin 1 PMID: 14684750
mutation analysis of the conserved regions in the ARAF gene in human colorectal adenocarcinoma PMID: 14688025
A-Raf residues are identified that bind to specific phosphoinositides, possibly as a mechanism to localize the enzyme to particular membrane microdomains rich in these phospholipids. PMID: 15736953
These data reveal that B-RAF is an important mediator of neuronal survival, migration, and dendrite formation and that A-RAF cannot fully compensate for these functions. PMID: 16980614
Positive regulation of A-RAF by phosphorylation of isoform-specific hinge segment and identification of novel phosphorylation sites. PMID: 18662992
A-Raf interacts with MEK1 and activates MEK1 by phosphorylation. PMID: 8621729

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Protein Families

Protein kinase superfamily, TKL Ser/Thr protein kinase family, RAF subfamily

Tissue Specificity

Predominantly in urogenital tissues.

Database Links

HGNC: 646

OMIM: 311010

KEGG: hsa:369

STRING: 9606.ENSP00000366244

UniGene: Hs.446641

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Antibody FAQs What Is an Antibody? How to Choose an Antibody for Scientific Research? The difference between primary antibody and secondary antibod The Meaning of Irregular Antibody Screening Tag Antibodies, Your Good Partner in Protein Research How to Choose the Loading Control Antibodies?

Related pathways

ErbB signaling pathway MAPK signaling pathway Regulation of actin cytoskeleton Vascular smooth muscle contraction Natural killer cell mediated cytotoxicity FoxO signaling pathway Insulin signaling pathway

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301-363-4651 (Available 9 a.m. to 5 p.m. CST from Monday to Friday)

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Address

No. 269, Shendun 5th Road, Donghu Hi-Tech Development Area, Hubei Province, 430206, P.R.China

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