Human 15 Lipoxygenase 2 Baculovirus-Insect cells Overexpression Lysate

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Human 15 Lipoxygenase 2 Baculovirus-Insect cells Overexpression Lysate: 产品信息

产品描述
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.
表达宿主
Baculovirus-Insect cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human ALOX15B (AAH35217.1)( Met1-Ile676) was expressed the N-terminal polyhistidine-tagged GST tag at the N-terminus.
分子量
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

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 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.
全称
arachidonate 15-lipoxygenase, type B
参考文献
  • Kilty I. et al., 2000, Eur J Biochem. 266 (1): 83-93.
  • Sigal E. et al., 1990, J Biol Chem. 265 (9): 5113-20.
  • Brash AR. et al., 1997, Proc Natl Acad Sci. 94 (12): 6148-52.
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