Western blot analysis of extracts from various cell lines using Arf6 Antibody.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Arf6 Antibody detects endogenous levels of total Arf6 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly165 of human Arf6 protein. Antibodies are purified by protein A and peptide affinity chromatography.
ADP-ribosylation factor (Arf) proteins are low molecular weight GTP binding proteins that belong to the Ras GTPase superfamily (1). Arf proteins are grouped into three distinct classes based on amino acid sequence and structural similarity, with Arf6 as the single class III protein to date. Arf6 is localized mainly to the plasma membrane and endosomes (1,2). This small GTPase interacts with PIP5K, PLD and Rac1, proteins important in lipid metabolism and actin regulation. Arf6 function depends upon its cycling between GDP- and GTP-bound states, which is regulated by associated GAP and GEF factors (3,4). Plasma membrane-associated Arf6 appears to play several functions during the many steps of membrane trafficking, including regulating membrane receptor internalization in both clathrin-dependent and independent pathways, endosomal recycling, and proximal actin reorganization and remodeling (5,6).
Explore pathways + proteins related to this product.
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