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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
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human ATP6V1A protein.
Eukaryotic cells contain ATP-driven proton pumps known as vacuolar H+-ATPases (v-ATPases) that acidify intracellular compartments and translocate protons across the plasma membrane (1,2). Intracellular v-ATPases play an important role in endocytosis and intracellular membrane trafficking, while plasma membrane v-ATPases are important in processes such as urinary acidification and bone resorption (1,2). Vacuolar ATPase enzymes are large, heteromultimeric protein complexes with component proteins found in either the V1 peripheral domain or the V0 integral domain (2). The cytoplasmic V1 domain contains a hexamer of A and B catalytic subunits, as well as a number of other protein subunits required for ATPase assembly and ATP hydrolysis. The integral V0 v-ATPase domain exhibits protein translocase activity and is responsible for transport of protons across the membrane (2). Research studies show that the v-ATPase subunits ATP6V0c, ATP6V0d1, ATP6V1A, ATP6V1B2, and ATP6V1D interact with the Ragulator protein complex and are essential for amino acid induced activation of mTORC1 on the surface of lysosomes (3). Additionally, ATP6V1A has been shown to interact with SARS-CoV-2 M protein and facilitate viral infection (4,5).
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