Member of the ARF family of GTPases within the superfamily of Ras-like GTPases. Similar to most GTPases, Arf1 cycles between GDP- and GTP-bound states; the GTP-bound form engages a set of diverse effectors to regulate cell processes. In particular, Arf1 regulates the formation of vesicle coats at different steps in the exocytic and endocytic pathways. 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). Arf1 N-terminal His6-tagged 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 1-181 of human Arf1.
- 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 1-181 of Arf1. The N-terminus retains an affinity tag: (His)6-Gly-Gly.
Calculated molecular weight: 21,690 g/mol.
Experimental molecular weight: NA.
Extension coefficient: 29,450 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.