|Datasheet||Specific References||Reviews||Related Products||Protocols|
|Baculovirus-Insect Cell lysate that Human USP46 transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).|
|A DNA sequence encoding the human USP46 [(Identical to the mouse USP46 (NP_808229.1)] (Met1-Glu366) was expressed and purified.|
|The secreted recombinant human USP46 consists of 477 amino acids and predicts a molecular mass of 55.2 KDa. The apparent molecular mass of the protein is approximately 64 Kda in SDS-PAGE under reducing conditions due to glycosylation.|
|Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.|
|Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.|
|12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.|
|Samples are stable for up to twelve months from date of receipt.|
|1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.|
|1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).|
|Store at 4℃. After re-dissolution, aliquot and store at -80℃.|
|Western blot (WB): Use at an assay dependent dilution.|
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
USP46 belongs to the peptidase C19 family, USP12/USP46 subfamily. Deubiquitinating enzymes (DUBs) are a large group of proteases which are also commonly referred to as deubiquitinating peptidases, deubiquitinating isopeptidases, deubiquitinases, ubiquitin proteases, ubiquitin hydrolyases, ubiquitin isopeptidases, or Dubs. They regulate ubiquitin-dependent metabolic pathways by cleaving ubiquitin-protein bonds. They also may act as negative and positive regulators of the ubiquitin system. Besides ubiquitin recycling, they are also involved in processing of ubiquitin precursors, in proofreading of protein ubiquitination and in disassembly of inhibitory ubiquitin chains. USP46 is a deubiquitinating enzyme that plays a role in behavior, possibly by regulating GABA action. It may act by mediating the deubiquitination of GAD1/GAD67. USP46 has almost no deubiquitinating activity by itself and requires the interaction with WDR48 to have a high activity and it is not involved in deubiquitination of monoubiquitinated FANCD2.