Human chymase 1 Baculovirus-Insect cells Overexpression Lysate


Human chymase 1 Baculovirus-Insect cells Overexpression Lysate: 产品信息

This Human chymase 1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of chymase 1 protein (Cat: 11482-H08B) from the overexpression lysate was verified.
Baculovirus-Insect cells
A DNA sequence encoding the human CMA1 (P23946) (Met 1-Asn 247) was fused with a polyhistidine tag at the C-terminus.
The secreted recombinant human CMA1 consists of 238 amino acids and predicts a molecular mass of 26.6 kDa. It migrates as an approximately 33 kDa band in SDS-PAGE under reducing conditions.

Human chymase 1 Baculovirus-Insect cells Overexpression Lysate: 使用指南

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.
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.
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.
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.
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human chymase 1 Baculovirus-Insect cells Overexpression Lysate: 别称

Human chymase Overexpression Lysate; Human CYH Overexpression Lysate; Human MCT1 Overexpression Lysate

chymase 1 背景信息

The STR polymorphism in the CMA1 gene is associated with asthma and that this association is even stronger with atopic asthma. CMA1 and IL-4 in atopic asthma and for IL-4 in atopy in general. The local angiotensin II system (LAS) has numerous functions, including the regulation of growth and differentiation in the gastrointestinal tract. Angiotensin II (AngII) may be generated by angiotensin-I-converting enzyme (ACE) or mast cell chymase (CMA1) and plays an important role in inflammatory processes, although opinions differ as to which AngII-generating enzyme is primarily associated with AngII-mediated effects. ACE in the gastric mucosa and the microvasculature of granulation tissue may represent a novel therapeutic target for the promotion of healing processes in gastritis and ulceration using ACE inhibitors or AT1R antagonists. The gene for mast cell chymase (CMA1) is an ideal candidate for investigating genetic predisposition to atopic asthma, as it is an important mediator of inflammation and remodeling in the asthmatic lung. (CMA1) is important for the generation of angiotensin II and therefore might be associated with the pathogenesis of hypertension.
chymase 1, mast cell
  • Takai S, et al. (2003) Effect of chymase inhibitor on vascular proliferation. Jpn J Pharmacol. 90 (3): 223-7.
  • Caughey GH, et al. (1993) The human mast cell chymase gene (CMA1): mapping to the cathepsin G/granzyme gene cluster and lineage-restricted expression. Genomics. 15 (3): 614-20.
  • Urata H, et al. (1991) Cloning of the gene and cDNA for human heart chymase. J Biol Chem. 266 (26): 17173-9.
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