Human, Mouse USP46 Baculovirus-Insect cells Overexpression Lysate

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Human, Mouse USP46 Baculovirus-Insect cells Overexpression Lysate: 产品信息

产品描述
This Human, Mouse USP46 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of USP46 protein (Cat: 12976-HNCB) from the overexpression lysate was verified.
表达宿主
Baculovirus-Insect cells
种属
Human, Mouse
蛋白构建信息
A DNA sequence encoding the human / mouse USP46 [(Identical to the mouse USP46 (NP_808229.1)] (Met1-Glu366) was expressed. Human and Mouse USP46 sequences are identical.
分子量
The secreted recombinant human / mouse USP46 consists of 366 amino acids and predicts a molecular mass of 42.4 KDa. The apparent molecular mass of the protein is approximately 43 Kda in SDS-PAGE under reducing conditions due to glycosylation.

Human, Mouse USP46 Baculovirus-Insect cells Overexpression Lysate: 使用指南

制备方法
Cell lysate was prepared by homogenization of the over-expressed cells 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.
裂解缓冲液
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.
使用建议
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.
缓冲液
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

USP46 背景信息

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 high activity and it is not involved in deubiquitination of monoubiquitinated FANCD2.
全称
ubiquitin specific peptidase 46
参考文献
  • Joo HY, et al. (2011) Regulation of histone H2A and H2B deubiquitination and Xenopus development by USP12 and USP46. J Biol Chem. 286(9):7190-201.
  • Kushima I, et al. (2010) Association study of ubiquitin-specific peptidase 46 (USP46) with bipolar disorder and schizophrenia in a Japanese population. J Hum Genet. 55(3):133-6.
  • Cohn MA, et al. (2009) UAF1 is a subunit of multiple deubiquitinating enzyme complexes. J Biol Chem. 284(8):5343-51.
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