Member of the ARF family of GTPases within the superfamily of Ras-like GTPases. Similar to most GTPases, Arf6 cycles between GDP- and GTP-bound states; the GTP-bound form engages a set of diverse effectors to regulate cell processes. In particular, Arf6 N-terminal His6-tagged regulates actin organization as well as endocytosis. Nucleotide cycling is regulated by guanine nucleotide exchange factors (GEFs), guanine dissociation inhibitors (GDIs) and GTPase activating proteins (GAPs).1,2
Protein is > 98% pure as determined by densitometry of the final sample submitted to SDS-PAGE and stained with Coomassie blue (Fig. 1). Arf6 was heterologously expressed in E. coli as a histidine-tagged protein and purified by a combination of affinity, ion exchange, and gel exclusion chromatographies. The histidine-tagged was not removed leaving a (His)6-Gly-Gly tag at the N-terminus followed by residues 1-175 of human Arf6.
- Donaldson JG, et al., Arf Family G Proteins and their regulators: roles in membrane transport, development and disease. Nat Rev Mol Cell Biol 12(6), 362-375 (2011).
- Wennerberg K, et al., The Ras superfamily at a glance. J Cell Sci 118, 843-846 (2005).
Species: Homo sapiens.
Sequence: Residues 1-175 of Arf6. The N-terminus retains an affinity tag: (His)6-Gly-Gly.
Calculated molecular weight: 21,076 g/mol.
Experimental molecular weight: NA.
Extension coefficient: 32,430 M-1cm-1 (assuming all cysteines remain reduced).
Amount: 100 or 500 μg.
Concentration: 1 mg/ml (determined by absorbance at 280 nm).
Buffer: 25 mM Tris pH 7.5, 150 mM NaCl, 5% glycerol and 1 mM DTT.
Form: Frozen liquid.
Storage and stability:
Storage: -80 °C. After initial thaw, aliquot and store at -80 °C until further use.
Stability: Stable for up to 12 months at -80 °C and up to 2 days at 4 °C. Repeated freeze-thaw cycles not recommended.
Notes: Please see accompanying sheet describing activity.