Human Lp-PLA2/PLA2G7 HEK293 Overexpression Lysate

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Human Lp-PLA2/PLA2G7 HEK293 Overexpression Lysate: 产品信息

产品描述
This Human Lp-PLA2/PLA2G7 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Lp-PLA2/PLA2G7 protein (Cat: 10848-H08H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human PLA2G7 (Q13093-1) precursor (Met 1-Asn 441) was expressed, with a C-terminal polyhistidine tag.
分子量
The secreted recombinant human PLA2G7 comprises 431 amino acids with a predicted molecular mass of 49.2 kDa. As a result of glycosylation, it migrates as an approximately 50-55 kDa band in SDS-PAGE under reducing conditions.

Human Lp-PLA2/PLA2G7 HEK293 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.

Human Lp-PLA2/PLA2G7 HEK293 Overexpression Lysate: 别称

Human LDL-PLA2 Overexpression Lysate; Human LP-PLA2 Overexpression Lysate; Human PAFAD Overexpression Lysate; Human PAFAH Overexpression Lysate

Lp-PLA2/PLA2G7 背景信息

Platelet-activating factor acetylhydrolase, also known as 1-alkyl-2-acetylglycerophosphocholine esterase, 2-acetyl-1-alkylglycero-phosphocholine esterase, Group-VIIA phospholipase A2, LDL-associated phospholipase A2, PAF 2-acylhydrolase, PLA2G7 and PAFAH, is a secreted protein that belongs to the AB hydrolase superfamily and Lipase family. PLA2G7 / PAFAH modulates the action of platelet-activating factor (PAF) by hydrolyzing the sn-2 ester bond to yield the biologically inactive lyso-PAF. It has specificity for substrates with a short residue at the sn-2 position. It is inactive against long-chain phospholipids. PLA2G7 / PAFAH is a potent pro- and anti-inflammatory molecule that has been implicated in multiple inflammatory disease processes, including cardiovascular disease. PLA2G7 also represents an important, potentially functional candidate in the pathophysiology of coronary artery disease (CAD). Defects in PLA2G7 are the cause of platelet-activating factor acetylhydrolase deficiency (PLA2G7 deficiency). It is a trait that is present in 27% of Japanese. It could have a significant physiologic effect in the presence of inflammatory bodily responses.
全称
phospholipase A2 group VII
参考文献
  • Stafforini D.M., et al., 1996, J. Clin. Invest. 97:2784-2791.
  • Yoshida H., et al., 1998, Thromb. Haemost. 80:372-375.
  • Yamada Y., et al., 1998, Metabolism 47:177-181.
  • Kruse S., et al., 2000, Am. J. Hum. Genet. 66:1522-1530. 
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