Human IFI30 HEK293 Overexpression Lysate

Price:
Size:
Number:

Human IFI30 HEK293 Overexpression Lysate: 产品信息

产品描述
This Human IFI30 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IFI30 protein (Cat: 12623-H08H) 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 IFI30 (P13284) (Met1-Lys243) with a C-terminal polyhistidine tag was expressed.
分子量
The recombinant human IFI30 is a disulfide-linked homodimer. The reduced monomer comprises 217 amino acids and has a predicted molecular mass of 24.7 kDa. The apparent molecular mass of the protein is approximately 31-34 kDa in SDS-PAGE under reducing conditions.

Human IFI30 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.

Human IFI30 HEK293 Overexpression Lysate: 别称

Human GILT Overexpression Lysate; Human IFI-30 Overexpression Lysate; Human IFI30 Overexpression Lysate; Human IP-30 Overexpression Lysate; Human IP30 Overexpression Lysate

IFI30 背景信息

IFI30 belongs to the GILT family. This family includes the two characterized human gamma-interferon-inducible lysosomal thiol reductase (GILT) sequences: P13284 and Q9UL08. It also contains several other eukaryotic putative proteins with similarity to GILT. The aligned region contains three conserved cysteine residues. Besides, the two GILT sequences possess a C-X(2)-C motif that is shared by some of the other sequences in the family. This motif is thought to be associated with disulfide bond reduction. IFI30 is a lysosomal thiol reductase that can reduce protein disulfide bonds. It facilitates the generation of MHC class II-restricted epitopes from disulfide bond-containing antigens by the endocytic reduction of disulfide bonds. It also facilitates MHC class I-restricted recognition of exogenous antigens containing disulfide bonds by CD8+ T-cells or cross-presentation. IFI30 may facilitate the complete unfolding of proteins destined for lysosomal degradation and plays an important role in antigen processing.
全称
interferon gamma inducible protein 30
参考文献
  • Haque MA, et al. (2002) Absence of gamma-Interferon-inducible Lysosomal Thiol Reductase in Melanomas Disrupts T Cell Recognition of Select Immunodominant Epitopes. J Exp Med. 195(10):1267-77.
  • Phan UT, et al. (2000) Gamma-interferon-inducible lysosomal thiol reductase (GILT). Maturation, activity, and mechanism of action. J Biol Chem. 275(34):25907-14.
  • Phan UT, et al. (2001) Multiple species express thiol oxidoreductases related to GILT. Immunogenetics. 53(4):342-6.
添加购物车成功! 添加购物车失败!请再次尝试 正在更新购物车,请稍后 U.S.A.