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KRAS (G12V), Isoform B, His-Tag, GppNHp-Loaded Recombinant

Recombinant human KRAS (GTPase), isoform b, encompassing amino acids 2-185. In this construct the N-terminal end of the protein contains a His-tag (6xHis) followed by an optimal TEV protease target sequence (TEV: tobacco etch virus cysteine protease #50308) . The protein also contains the cancer-associated mutation G12V. The protein was affinity purified and loaded with GppNHp, a non-hydrolyzable GTP analog. Unbound GppNHp was removed by spin column.

Product Specifications

Background

RAS is a group of small GTPase proteins encoded by 3 genes (HRAS, NRAS, and KRAS), which play critical roles in regulating cell proliferation. KRAS mutations are responsible for more than 30% of human cancers. The G12V mutation favors the activated (GTP-bound) state of KRAS, amplifying signaling pathways that lead to oncogenesis. Recent studies have led to the discovery of a small molecule called AMG510 (Amgen) that locks KRAS conformation in the GDP-bound (inactive) state, thereby blocking KRAS (G12V) -mediated signaling pathway.The KRAS (Kirsten rat sarcoma virus) gene is subject to alternative splicing, resulting in two isoforms: KRAS-A and KRAS-B. These isoforms differ by amino acids 151,153,165, and 166 and within the hypervariable region (amino acids 167-189) . KRAS-B contains a long polybasic stretch, while KRAS-A has a shorter polybasic region with a palmitoylation site. These differences confer distinct biological characteristics to the two isoforms. When studying a mutant of KRAS, it is important to know which isoform is being studied to make sure that the correct wild-type isoform is used for comparison. The identification of new strategies targeting KRAS G12V will bring large benefits in cancer therapy.

UniProt

P01116

Tag

N-terminal His-tag

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Purity

≥ 90%

Format

Aqueous buffer solution

Shipping Conditions

-80°C

Storage Conditions

At least 6 months at -80°C.

Calculated Molecular Weight

22 kDa

Formulation

20 mM HEPES, pH 7.4,150 mM NaCl, and 1 mM DTT

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