Mouse IgG1 HEK293 Overexpression Lysate

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Mouse IgG1 HEK293 Overexpression Lysate: 产品信息

产品描述
This Mouse IgG1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IgG1 protein (Cat: 10690-MNAH) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Mouse
蛋白构建信息
The Fc region of mouse IgG1 (P01868-1) (Val 98-Lys 324) (one aa mutation, 102 Cys / Ser) was expressed and purified.
分子量
The recombinant mouse IgG1 Fc consists of 227 amino acids and has a predicted molecular mass of 25.8 kDa. As a result of glycosylation, the apparent molecular mass of rmFc is approximately 32 kDa in SDS-PAGE under reducing conditions.

Mouse IgG1 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.

Mouse IgG1 HEK293 Overexpression Lysate: 别称

Mouse IgG1 Overexpression Lysate; Mouse Igh-4 Overexpression Lysate; Mouse VH7183 Overexpression Lysate

IgG1 背景信息

As a monomeric immunoglobulin that is predominately involved in the secondary antibody response and the only isotype that can pass through the human placenta, Immunoglobulin G (IgG) is synthesized and secreted by plasma B cells, and constitutes 75% of serum immunoglobulins in humans. IgG antibodies protect the body against the pathogens by agglutination and immobilization, complement activation, toxin neutralization, as well as the antibody-dependent cell-mediated cytotoxicity (ADCC). IgG tetramer contains two heavy chains (5 kDa ) and two light chains (25 kDa) linked by disulfide bonds, that is the two identical halves form the Y-like shape. IgG is digested by pepsin proteolysis into Fab fragment (antigen-binding fragment) and Fc fragment ("crystallizable" fragment). IgG1 is most abundant in serum among the four IgG subclasses (IgG1, 2, 3 and 4) and binds to Fc receptors (FcγR ) on phagocytic cells with high affinity. Fc fragment is demonstrated to mediate phagocytosis, trigger inflammation, and target Ig to particular tissues. Protein G or Protein A on the surface of certain Staphylococcal and Streptococcal strains specifically binds with the Fc region of IgGs, and has numerous applications in biotechnology as a reagent for affinity purification. Recombinant IgG Fc Region is suggested to represent a potential anti-inflammatory drug for treatment of human autoimmune diseases.
参考文献
  • Carosella ED. et al., 1988, Cell Immunol. 112: 262-70.
  • Eckle I. et al., 1990, Clin Exp Immunol. 81: 352-6.
  • Lorriaine C. et al., 1995, J Biol Chem. 270:8164-71.
  • Gomez-Guerrero C. et al., 2000, J Immunol. 164: 2092-101.
  • Sprague ER. et al., 2004, J Biol Chem. 14: 14184-93.
  • Anthony RM. et al., 2008, Science. 320: 373-6.
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