Rat CD64 HEK293 Overexpression Lysate

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Rat CD64 HEK293 Overexpression Lysate: 产品信息

产品描述
This Rat CD64 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD64 protein (Cat: 80016-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 FCGR1A (NP_001094306.1) extracellular domain (Met 1-Pro 285) was expressed, fused with a polyhistidine tag at the C-terminus.
分子量
The recombinant rat FCGR1A comprises 280 amino acids and predicts a molecular mass of 31.6 kDa. The apparent molecular mass of the rat FCGR1A is approximately 40-42 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Rat CD64 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.

CD64 背景信息

High affinity immunoglobulin gamma Fc receptor I, also known as FCGR1 and CD64, is an integral membraneglycoprotein and a member of the immunoglobulin superfamily. CD64 is a high affinity receptor for the Fc region of IgG gamma and functions in both innate and adaptive immune responses. Receptors that recognize the Fc portion of IgG function in the regulation of immune response and are divided into three classes designated CD64, CD32, and CD16. CD64 is structurally composed of asignal peptidethat allows its transport to the surface of a cell, threeextracellularimmunoglobulin domainsof the C2-type that it uses to bind antibody, a hydrophobictransmembrane domain, and a short cytoplasmic tail. CD64 isconstitutivelyfound on only macrophages and monocytes, but treatment of polymorphonuclear leukocyteswith cytokines likeIFNγandG-CSFcan induce CD64 expression on these cells. The inactivation of the mouse CD64 resulted in a wide range of defects in antibody Fc-dependent functions. Mouse CD64 is an early participant in Fc-dependent cell activation and in the development of immune responses.
全称
Fc receptor, IgG, high affinity I
研究领域
参考文献
  • Allen J.M., et al.,(1988), Nucleotide sequence of three cDNAs for the human high affinity Fc receptor (FcRI). Nucleic Acids Res. 16:11824-11824.
  • Allen J.M., et al., (1989), Isolation and expression of functional high-affinity Fc receptor complementary DNAs.Science 243:378-381.
  • Porges A.J., et al.,(1992), Novel Fc gamma receptor I family gene products in human mononuclear cells.J. Clin. Invest. 90:2102-2109.
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