Product Name:
KRas-pS181
Product Number:
ab-pn862
Target Full Name: GTPase Kirsten Sarcoma Protein Ras
Target Alias: GTPase HRas; GTPase KRas; GTPase NRas; Ha Ras; K Ras; K ras p21; KRAS; NRAS; p21ras; RASH2
Product Type Specific: K-Ras G protein phosphosite-specific antibody
Antibody Code: PN862
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S181
Protein UniProt: P01116
Protein SigNET: KRas
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human KRas
Antibody Immunogen Sequence: KKK(pS)KTK(βA)C
Antibody Immunogen Description: Corresponds to amino acid residues K178 to K184. Phosphorylation of K-Ras isoform 2 at S181 stimulates its activity and its interaction with RhoA, and inhibits binding to calmodulin, PDE6D and Raf1. Phosphorylation of this site has not been recorded by high throughput mass spectrometry studies in PhosphoSitePlus. KRAS is known to be phosphorylated at this site in vitro by PKACA, and PKCa.
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: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Mouse.
Related Product 1: KRAS expression antibody (Cat. No.: AB-NN281-2)
Related Product 2: Ras expression antibody (Cat. No.: AB-NN281-1)
Scientific Background: The 21 kDa guanine-nucleotide binding proteins (K-Ras, H-Ras and N-Ras) cycle between active (GTP-bound) and inactive (GDP-bound) forms (1). Receptor tyrosine kinases and G protein-coupled receptors activate Ras, which then stimulates the Raf-MEK-MAPK pathway (2-4). GTPase-activating proteins (GAP) normally facilitate the inactivation of Ras. However, in 30% of human tumours, point mutations in Ras prevent the GAP-mediated inhibition of this pathway (5). The most common oncogenic Ras mutation found in tumours is Gly-12 to Asp (G12D), which prevents Ras inactivation, possibly by increasing the overall rigidity of the protein (5,6). H-Ras protein (Harvey rat sarcoma viral oncogene homologue) is associated with the inner plasma membrane, often via a cycle of palmitoylation that facilitates its movement between the plasma membrane and the Golgi apparatus. Activated H-Ras recruits and stimulates downstream signaling proteins, including c-Raf and PI 3-kinase, which propagate signals for cellular proliferation. Activating mutations in the HRAS gene are found in various human cancers, including bladder cancer, follicular thyroid cancer, and oral squamous cell carcinoma. These mutations commonly impair the protein's GTPase activity, locking it in the active state and causing uncontrolled cell growth. Somatic mutations in HRAS are associated with Costello syndrome, a rare disorder characterized by prenatally increased growth, postnatal deficiency, and tumour predisposition. This description may include information annotated by UniProt and/or Google AI.