Human APOL1 Baculovirus-Insect cells Overexpression Lysate

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Human APOL1 Baculovirus-Insect cells Overexpression Lysate: 产品信息

产品描述
This Human APOL1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of APOL1 protein (Cat: 13910-H08B) 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 APOL1 (Met 1-Leu398) (Q2KHQ6) was expressed, with a C-terminal polyhistidine tag.
分子量
The secreted recombinant human APOL1 consists of 381 amino acids and predicts a molecular mass of 42.5 KDa. The apparent molecular mass of the protein is approximately 44 KDa in SDS-PAGE under reducing conditions due to glycosylation.

Human APOL1 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 APOL1 Baculovirus-Insect cells Overexpression Lysate: 别称

Human APO-L Overexpression Lysate; Human APOL Overexpression Lysate; Human APOL-I Overexpression Lysate; Human APOL1 Overexpression Lysate; Human FSGS4 Overexpression Lysate

APOL1 背景信息

APOL1, also known as apolipoprotein L1, is a minor apoprotein component of HDL (High-density lipoprotein) or 'good cholesterol' which is synthesized in the liver and also in many other tissues, including pancreas, kidney, and brain. APOL1 belongs to the apolipoprotein L family. It may play a role in lipid exchange and transport throughout the body. It may also participate in reverse cholesterol transport from peripheral cells to the liver. Defects in APOL1 are the cause of focal segmental glomerulosclerosis type 4 (FSGS4). It is a renal pathology defined by the presence of segmental sclerosis in glomeruli and resulting in proteinuria, reduced glomerular filtration rate and edema. Renal insufficiency often progresses to end-stage renal disease, a highly morbid state requiring either dialysis therapy or kidney transplantation.
全称
apolipoprotein L1
参考文献
  • Genovese G, et al. (2010) Association of Trypanolytic ApoL1 Variants with Kidney Disease in African-Americans. Science. 329 (5993): 841-5.
  • Tzur S, et al. (2010) Missense mutations in the APOL1 gene are highly associated with end stage kidney disease risk previously attributed to the MYH9 gene. Human Genetics 128 (3): 345-50.
  • Hu CA, et al. (2012) Human apolipoprotein L1 (ApoL1) in cancer and chronic kidney disease. FEBS Lett. 586 (7): 947-55.
  • Papeta N, et al. (2011) APOL1 variants increase risk for FSGS and HIVAN but not IgA nephropathy. J Am Soc Nephrol. 22 (11): 1991-6.
  • Fine DM, et al. (2012) APOL1 risk variants predict histopathology and progression to ESRD in HIV-related kidney disease. J Am Soc Nephrol. 23 (2): 343-50.
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