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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: 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: 10
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human TSG101 protein.
Tumor susceptibility gene 101 protein (TSG101) is a multi-domain cytoplasmic protein and fundamental component of ESCRT complex I, which is intimately involved in regulating the trafficking of proteins throughout the endosomal compartment (1). In addition to regulating the sorting of ubiquitinated cargo proteins within the endosomal compartment, TSG101 is involved in regulating diverse biological processes, such as cell proliferation, viral budding and release, and exosome biosynthesis (2-6). While it is clear that TSG101 expression levels are tightly controlled and complete loss of expression causes embryonic lethality due to cell cycle arrest and cell death, the role of TSG101 in malignancy has yet to be fully elucidated (7). Indeed, TSG101 was initially dubbed a tumor suppressor but there are multiple studies that demonstrate an elevated expression in human cancers (8-10).
UniProt ID: Q99816
Entrez-Gene Id: 7251
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