Human Lp-PLA2/PLA2G7 HEK293 Overexpression Lysate: Product Information
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.
A DNA sequence encoding the human PLA2G7 (Q13093-1) precursor (Met 1-Asn 441) was expressed, with a C-terminal polyhistidine tag.
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: Usage Guide
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.
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).
Stability & Storage
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 Lp-PLA2/PLA2G7 HEK293 Overexpression Lysate: Alternative Names
Human LDL-PLA2 Overexpression Lysate; Human LP-PLA2 Overexpression Lysate; Human PAFAD Overexpression Lysate; Human PAFAH Overexpression Lysate
Lp-PLA2/PLA2G7 Background Information
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 secreted protein which belongs to theAB 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 a 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 which is present in 27% of Japanese. It could have a significant physiologic effect in the presence of inflammatory bodily responses.
phospholipase A2 group VII
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