Human PPT1 HEK293 Overexpression Lysate

Price:
Size:
Number:

Human PPT1 HEK293 Overexpression Lysate: 产品信息

产品描述
This Human PPT1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PPT1 protein (Cat: 14703-H02H) 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 PPT1 (P50897-1)(Met1-Gly306) was expressed with the Fc region of human IgG1 at the C-terminus.
分子量
The recombinant human PPT1/Fc comprises 520 amino acids and has a predicted molecular mass of 58.3 kDa. The apparent molecular mass of the protein is approximately 58-67 kDa in SDS-PAGE under reducing conditions.

Human PPT1 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 PPT1 HEK293 Overexpression Lysate: 别称

Human CLN1 Overexpression Lysate; Human INCL Overexpression Lysate; Human PPT Overexpression Lysate; Human PPT1 Overexpression Lysate

PPT1 背景信息

Mutations in the depalmitoylating enzyme gene, PPT1, cause the infantile form of Neuronal Ceroid Lipofuscinosis (NCL), an early onset neurodegenerative disease. Mutations in palmitoyl protein thioesterase-1 (PPT1) have been found to cause the infantile form of neuronal ceroid lipofuscinosis, which is a lysosomal storage disorder characterized by impaired degradation of fatty acid-modified proteins with accumulation of amorphous granular deposits in cortical neurons, leading to mental retardation and death. PPT1 catalyzes the cleavage of thioester linkages in S-acylated (palmitoylated) proteins and its deficiency leads to abnormal accumulation of thioesterified polypeptides (ceroid) in lysosomes causing INCL pathogenesis.
全称
palmitoyl-protein thioesterase 1
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.