Human 15 Lipoxygenase 2 Baculovirus-Insect cells Overexpression Lysate: 产品信息
This Human 15 Lipoxygenase 2 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of 15 Lipoxygenase 2 protein (Cat: 14564-H20B) from the overexpression lysate was verified.
A DNA sequence encoding the human ALOX15B (AAH35217.1)( Met1-Ile676) was expressed the N-terminal polyhistidine-tagged GST tag at the N-terminus.
The secreted recombinant human ALOX15B consists of 913 amino acids and predicts a molecular mass of 103.7 KDa. The apparent molecular mass of the protein is approximately 93 KDa in SDS-PAGE under reducing conditions due to glycosylation.
Human 15 Lipoxygenase 2 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.
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 15 Lipoxygenase 2 Baculovirus-Insect cells Overexpression Lysate: 别称
Human 15-LOX-2 Overexpression Lysate; Human ALOX15B Overexpression Lysate
15 Lipoxygenase 2 背景信息
ALOX15B is a member of the lipoxygenase family of structurally related nonheme iron dioxygenases involved in the production of fatty acid hydroperoxides. ALOX15B converts arachidonic acid exclusively to 15S-hydroperoxyeicosatetraenoic acid, while metabolizing linoleic acid less effectively. ALOX15B gene is located in a cluster of related genes and a pseudogene that spans approximately 1 kilobases on the short arm of chromosome 17. Alternatively spliced transcript variants encoding different isoforms have been described.
arachidonate 15-lipoxygenase, type B
Kilty I. et al., 2000, Eur J Biochem. 266 (1): 83-93.
Sigal E. et al., 1990, J Biol Chem. 265 (9): 5113-20.