Human Doppel / PRND HEK293 Overexpression Lysate: Product Information
This Human Doppel / PRND overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Doppel / PRND protein (Cat: 15113-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human PRND (NP_036541.2) (Met1-Gly152) was expressed with a polyhistidine tag at the C-terminus.
The recombinant human PRND consists 137 amino acids and predicts a molecular mass of 15.9 kDa.
Human Doppel / PRND HEK293 Overexpression Lysate: Usage Guide
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).
Stability & Storage
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 Doppel / PRND HEK293 Overexpression Lysate: Alternative Names
Human dJ1068H6.4 Overexpression Lysate; Human DOPPEL Overexpression Lysate; Human DPL Overexpression Lysate; Human PrPLP Overexpression Lysate
Doppel / PRND Background Information
Doppel (PRND) is a paralogue of the mammalian prion (PRNP) gene. It is abundant in testis and, unlike PRNP, it is expressed at low levels in the adult central nervous system (CNS). Besides, doppel overexpression correlates with some prion-disease pathological features, such as ataxia and death of cerebellar neurons.Prion-related protein doppel gene (PRND), as an essential member of the mammalian prion gene family, is associated with the scrapie susceptibility as well as phenotype traits, so the genetic variation of the PRND has been highly concerned recently, including the single nucleiotide polymorphism (SNP) and insertion/deletion (indel).
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