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.
A DNA sequence encoding the human C5orf15 (NP_064584.1) (Met1-Asp196) was expressed with a C-terminal polyhistidine tag.
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
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.
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.
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.
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
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.
chromosome 5 open reading frame 15
Bonkobara M., et al.,(2003), Identification of novel genes for secreted and membrane-anchored proteins in human keratinocytes. Br. J. Dermatol. 148:654-664.