Product Name: ARaf-2
Product Number: AB-NK205-3
Size: 25 µg      Price:89.00
      $US
Target Full Name: A-Raf proto-oncogene serine/threonine-protein kinase

Target Alias: ARaf; A-RAF; ARaf1; A-raf-1; Kinase A-Raf; PKS; PKS2; Proto-oncogene Pks; V-raf murine sarcoma 3611 viral oncogene; RP1-230G1.1-005; RP1-230G1_1; A6NIT1; B4DMG5; Q5H9B2; Q96II5ENSG00000078061

Product Type Specific: Protein kinase pan-specific antibody

Antibody Code: NK205-3

Antibody Target Type: Pan-specific

Protein UniProt: P10398

Protein SigNET: P10398

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human Raf-A (ARaf) sequence peptide Cat. No.: PE-01ASF80

Antibody Immunogen Sequence: CGRKTVTAWDTAIAPL

Antibody Immunogen Description: Corresponds to amino acid residues G67 to L81

Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.

Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.

Antibody Concentration: 1 mg/ml

Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol

Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.

Product Use: Western blotting | Antibody microarray

Antibody Dilution Recommended: 1 µg/ml for immunoblotting

Antibody Species Reactivity: Human; Mouse

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 63-70 kDa.

Antibody Specificity: High

Antibody Cross Reactivity: No immunoreactivity on protein dot blots with recombinant human B-Raf and Raf1.

Related Product 1: ARaf-2 blocking peptide

Related Product 2: A-Raf-pY302 phosphosite-specific antibody (Cat. No.: AB-PK500)

Related Product 3: ARaf-1 pan-specific antibody (Cat. No.: AB-NK205-5)

Related Product 4: ARaf-3 pan-specific antibody (Cat. No.: AB-NK205-4)

Scientific Background: A-Raf (RafA) is a protein-serine/threonine kinase of the TKL group and RAF family. It is involved in the transduction of mitogenic signals from the cell membrane receptors via Ras and MEK1/2 to the nucleus. It has been implicated in playing a role in the postsynaptic responses of hippocampal neuron. Phosphorylation of S214, S257, S262, S264, Y301, Y302, S432, T442, T452, T455 and S582 increases phosphotransferase activity. It is inhibited by Nexavar/Sorafenib (IC50>50 nM). A-Raf may be an oncoprotein (OP) based on its similarity to Raf1 and B-Raf. However, it does not display a higher than normal rate of mutation than most proteins in human tumours. The active form of the protein kinase acts to promote tumour cell proliferation.
Figure 1. Dot blotting ARaf-2 antibody with recombinant purified proteins.


Figure 2. Immunoblotting of various cell lines with AB-NK205-1 antibody at a 250 X dilution. The target protein A-Raf is indicated. The expected molecular mass of A-Raf is 67.6 kDa. An alternatively spliced 20.9 kDa isoform is also produced from the first third of the full length sequence. Each lane was loaded with 20 µg of cell lysate protein.