For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 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.
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: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
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: 263
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val31 of human VAMP8 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Proteins in the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex are integral membrane proteins involved in vesicle transport and membrane fusion by pairing of vesicular SNAREs (v-SNAREs) with cognate target SNAREs (t-SNAREs) (reviewed in 1,2). Vesicle associated membrane protein 8 (VAMP8), also known as endobrevin, is a v-SNARE originally found preferentially localized to early endosomes (3). VAMP8 knockout mice did not show abnormal endosomal vesicular trafficking, perhaps having a redundant role with other VAMP family members (4). Instead, research studies have shown that VAMP8 is widely expressed in exocrine tissues and has a critical role in the exocytosis pathways of a variety of cells (4-9). In addition, lysosome localized VAMP8 has been shown to play a role in autophagosome/lysosome fusion during antimicrobial (xenophagy) and canonical starvation induced autophagy (5).
UniProt ID: Q9BV40
Entrez-Gene Id: 8673
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