This Mouse PLA2G1B overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PLA2G1B protein (Cat: 50329-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse PLA2G1B (NP_035237.1) extracellular domain (Met 1-Cys 146) was fused with a polyhistidine tag at the C-terminus.
The secreted recombinant mouse PLA2G1B consists of 142 amino acids and has a predicted molecular mass of 16.3 kDa. rmPLA2G1B migrates as an approximately 18 kDa band in SDS-PAGE under reducing conditions.
Mouse PLA2G1B HEK293 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.
phospholipase A2, also known as Phosphatidylcholine 2-acylhydrolase 1B, Group IB phospholipase A2, PLA2 and PLA2G1B, is a secreted protein that belongs to the phospholipase A2 family. Phospholipase A2 / PLA2G1B catalyzes the release of fatty acids from glycero-3-phosphocholines. The best known varieties are the digestive enzymes secreted as zymogens by the pancreas of mammals. Sequences of pancreatic Phospholipase A2 / PLA2G1B enzymes from a variety of mammals have been reported. One striking feature of these enzymes is their close homology to venom phospholipases of snakes. Other forms of Phospholipase A2 / PLA2G1B have been isolated from brain, liver, lung, spleen, intestine, macrophages, leukocytes, erythrocytes, inflammatory exudates, chondrocytes, and platelets. Mice lacking in Phospholipase A2 / PLA2G1B are resistant to obesity and diabetes induced by feeding a diabetogenic high-fat/high-carbohydrate diet. Oral supplementation of a diabetogenic diet with the PLA2G1B inhibitor methyl indoxam effectively suppresses diet-induced obesity and diabetes. PLA2G1B inhibition may be a potentially effective oral therapeutic option for treatment of obesity and diabetes.
phospholipase A2, group IB (pancreas)
Labonté,E.D. et al., 2006, Diabetes. 55 (4) :935-41.
Mounier,C.M. et al.,2008, Br J Cancer. 98 (3):587-95.
Hui,D.Y. et al., 2009, Br J Pharmacol. 157 (7):1263-9.
Labonté,E.D. et al., 2010, FASEB J. 24 (7):2516-24.