Rat Beta-2 microglobulin HEK293 Overexpression Lysate

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Rat Beta-2 microglobulin HEK293 Overexpression Lysate: 产品信息

产品描述
This Rat Beta-2 microglobulin overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Beta-2 microglobulin protein (Cat: 80423-R08H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Rat
蛋白构建信息
A DNA sequence encoding the rat B2M (P07151) (Met 1-Met 119) was expressed, fused with a polyhistidine tag at the C-terminus.
分子量
The secreted recombinant rat B2M comprises 110 amino acids and predicts a molecular mass of 13 kDa. The apparent molecular mass of the rat B2M is approximately 15 kDa in SDS-PAGE under reducing conditions.

Rat Beta-2 microglobulin HEK293 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.

Beta-2 microglobulin 背景信息

B2M, also known as β2-Microglobulin or CDABP0092, is a component of MHC class I molecules found expression in all nucleated cells (excludes red blood cells). The major function of MHC class I moleculesis is to display fragments of proteins from within the cell to T-cells and cells containing foreign proteins will be attacked. B2M (β2-Microglobulin) is a low molecular weight protein. It was demonstrated that B2M (β2-Microglobulin) was localized in the membranes of nucleated cells and was found to be associated with HL-A antigens. B2M (β2- Microglobulin) is present in free form in various body fluids and as a subunit of histocompatibility antigens on cell surfaces lateral to the α3 chain. Unlike α3, β2 has no transmembrane region. Directly above β2 lies the α1 chain, which itself is lateral to the α2. In the absence of B2M (β2 microglobulin), very limited amounts of MHC class I (classical and non-classical) molecules can be detected on the surface. In the absence of MHC class I, CD8 T cells, a subset of T cells involved in the development of acquired immunity cannot develop. Low levels of B2M (β2 microglobulin) can indicate non-progression of HIV.
全称
beta-2 microglobulin
参考文献
  • Poulik MD, et al. (1979) Beta 2-Microglobulin: methods and clinical applications. CRC Ctit Rev Clin Lab Sci. 10(3): 225-45.
  • Poulik MD, et al. (1975) Beta2-Microglobulins. Contemp Top Mol Immunol. 4: 157-204.
  • Berggard I. (1976) Beta2-Microglobulins: isolation, properties, and distribution. Fed Proc. 35(5): 1167-70.
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