Human LAG3 HEK293 Overexpression Lysate

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Human LAG3 HEK293 Overexpression Lysate: 产品信息

产品描述
This Human LAG3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of LAG3 protein (Cat: HPLC-16498-HNAH) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human LAG3 (P18627) (Met1-Arg440) was expressed with an initial Met.
分子量
The recombinant human LAG3 consists 418 amino acids and predicts a molecular mass of 45.3 kDa.

Human LAG3 HEK293 Overexpression Lysate: 使用指南

制备方法
Cell lysate was prepared by homogenization 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. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
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 LAG3 HEK293 Overexpression Lysate: 别称

Human CD223 Overexpression Lysate; Human LAG-3 Overexpression Lysate

LAG3 背景信息

LAG3 (Lymphocyte Activating 3) is a Protein Coding gene. 2 alternatively spliced human isoforms have been reported. LAG3, also known as CD223 and Lymphocyte activation gene 3, belongs to the immunoglobulin (Ig) superfamily. The LAG3 gene contains 8 exons. It is selectively expressed in activated T and NK cells. LAG3 contains 4 extracellular Ig-like domains and has a negative regulatory function in T cells. It also acts as a new marker of T cell-induced B cell activation. As a soluble molecule, LAG3 activates antigen-presenting cells through MHC class II signaling, leading to increased antigen-specific T-cell responses in vivo. Diseases associated with LAG3 include Smoldering Myeloma and Kyphoscoliotic Heart Disease.
全称
lymphocyte-activation gene 3
研究领域
参考文献
  • Sigrid Hannier. et al., 1998, The Journal of Immunology. 161(8): 4058-65.
  • Triebel F. et al., 1990, J Exp Med. 171 (5): 1393-405.
  • Baixeras E. et al., 1992, J Exp Med. 176 (2): 327-37.
  • Huard B. et al., 1997, Proc Natl Acad Sci. 94 (11): 5744-9.
  • Kisielow M. et al., 2005, Eur J Immunol. 35 (7): 2081-8.
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