Human p67phox Baculovirus-Insect cells Overexpression Lysate

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Human p67phox Baculovirus-Insect cells Overexpression Lysate: 产品信息

产品描述
This Human p67phox overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of p67phox protein (Cat: 15669-HNCB) 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 NCF2 (AAH01606.1) (Met1-Val526) was expressed and purified with two additional amino acids (Gly & Pro) at the N-terminus.
分子量
The secreted recombinant human NCF2 consists of 528 amino acids and predicts a molecular mass of 59.9 KDa. The apparent molecular mass of the protein is approximately 60 KDa in SDS-PAGE under reducing conditions due to glycosylation.

Human p67phox 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 p67phox Baculovirus-Insect cells Overexpression Lysate: 别称

Human NCF-2 Overexpression Lysate; Human NOXA2 Overexpression Lysate; Human P67-PHOX Overexpression Lysate; Human P67PHOX Overexpression Lysate

p67phox 背景信息

NCF2 (Neutrophil Cytosolic Factor 2, also known as NCF-2 and p67phox) is a Protein Coding gene. 4 alternatively spliced human isoforms have been reported. This gene encodes neutrophil cytosolic factor 2, the 67-kilodalton cytosolic subunit of the multi-protein NADPH oxidase complex found in neutrophils. This oxidase produces a burst of superoxide which is delivered to the lumen of the neutrophil phagosome. NCF2 belongs to the NCF2/NOXA1 family. NCF2, NCF1, and a membrane-bound cytochrome b558 are required for activation of the latent NADPH oxidase. Mutations in the NCF2 gene, as well as in other NADPH oxidase subunits, can result in chronic granulomatous disease, a disease that causes recurrent infections by catalase-positive organisms.
全称
neutrophil cytosolic factor 2
参考文献
  • Wientjes FB. et al., 1996, Semin Cell Biol. 6 (6): 357-65.
  • DeLeo FR. et al., 1997, J Leukoc Biol. 60 (6): 677-91.
  • Dorseuil O. et al., 1997, C R Seances Soc Biol Fil. 191 (2): 237-46.
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