Human KCT2 HEK293 Overexpression Lysate

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Human KCT2 HEK293 Overexpression Lysate: 产品信息

产品描述
This Human KCT2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of KCT2 protein (Cat: 13794-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 C5orf15 (NP_064584.1) (Met1-Asp196) was expressed with a C-terminal polyhistidine tag.
分子量
The recombinant human C5orf15 comprises 158 amino acids and has a predicted molecular mass of 17.5 kDa. The apparent molecular mass of the protein is approximately 33-47 kDa in SDS-PAGE under reducing conditions.

Human KCT2 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 KCT2 HEK293 Overexpression Lysate: 别称

Human HTGN29 Overexpression Lysate; Human KCT2 Overexpression Lysate

KCT2 背景信息

C5orf15 (Chromosome 5 Open Reading Frame 15, also known as KCT2) is a Protein Coding gene. KCT2 gene maps to human chromosome 5q31.1 and is conserved in human, chimpanzee, cow, rat, and chicken. KCT2 is a 265 amino acid single-pass type I membrane protein that is widely expressed in the thyroid, kidney, and other tissues. Chromosome 5 is associated with Cockayne syndrome through the ERCC8 gene and familial adenomatous polyposis through the adenomatous polyposis coli (APC) tumor suppressor gene. Chromosome 5 contains 181 million base pairs and comprises nearly 6% of the human genome. Diseases associated with C5orf15 include Arrhythmogenic Right Ventricular Dysplasia, Familial, 5, and Amyotrophic Lateral Sclerosis 11.
全称
chromosome 5 open reading frame 15
参考文献
  • Bonkobara M., et al.,(2003), Identification of novel genes for secreted and membrane-anchored proteins in human keratinocytes. Br. J. Dermatol. 148:654-664.
  • Yu W, et al., (1997), Large-scale concatenation cDNA sequencing. Genome Res. 7(4):353-8.
  • Ota T., et al.,(2004), Complete sequencing and characterization of 21,243 full-length human cDNAs.Nat. Genet. 36:40-45.
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