Product Name: Raf-A-pY302
Product Number: AB-PK500
Size: 25 µg      Price:89.00
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 phosphosite-specific antibody

Antibody Code: PK500

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: Y302

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-04AFW99

Antibody Immunogen Sequence: SGY(pY)WEV(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues S299 to V305; Just before kinase catalytic domain

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.This antibody was also subject to negative purification over phosphotyrosine-agarose.

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: 2 µg/ml for immunoblotting

Antibody Potency: Strong immunoreactivity with immunogen peptide on dot blots.

Antibody Species Reactivity: Human

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: Very high

Antibody Cross Reactivity: No significant cross-reactive proteins detected in HeLa, Jurkat and seastar oocytes.

Related Product 1: A-Raf-pY302 blocking peptide

Related Product 2: Raf-A-1 pan-specific antibody (Cat. No.: AB-NK205-1)

Related Product 3: Raf-A-2 pan-specific antibody (Cat. No.: AB-NK205-3P)

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

Related Product 5: Raf-A-CT (RafA-4) pan-specific antibody (Cat. No.: AB-NK205)

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. Epitope mapping of A-Raf-pY302 antibody with similar phosphopeptides on dot blots.

Figure 2. Identification of phosphosites related to A-Raf-pY302.