PrPC Overexpression Lysate

Western Blot: Prion protein Overexpression Lysate (Adult Normal) [NBL1-14799] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Prion protein.

Product Discontinued

PrPC Overexpression Lysate Summary

• Description: Transient overexpression lysate of prion protein (PRNP), transcript variant 2 The lysate was created in HEK293T cells, using Plasmid ID RC216132 and based on accession number NM_183079. The protein contains a C-MYC/DDK Tag. You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: PRNP

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 25.2 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

Alternate Names for PrPC Overexpression Lysate

• CD230
• CJD
• fatal familial insomnia)
• GSS
• prion protein (p27-30)
• prion protein PrP
• prion protein
• prion-related protein
• PRIPMGC26679

Background

Prion diseases are transmissible neurodegenerative disorders which affect a range of mammalian species. In humans they can be inherited and sporadic as well as acquired by exposure to human prions. Prions appear to be composed principally of a conformational isomer of host-encoded prion protein and propagate by recruitment of cellular prion protein (1). The function of the cellular prion protein (PrP) is still poorly understood. It has been proposed that one unprecedented role for PrP is against Bax-mediated neuronal apoptosis. It has been shown that PrP potently inhibits Bax-induced cell death in human primary neurons (2). An impaired synaptic inhibition may be involved in the epileptiform activity seen in Creutzfeldt-Jakob and other neurodegenerative diseases and it is believed that loss of function of PrP may contribute to the early synaptic loss and neuronal degeneration seen in these diseases (3).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: PRNP