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PERK Overexpression Lysate

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Western Blot: PERK Overexpression Lysate (Adult Normal) [NBL1-10177] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PERK.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

PERK Overexpression Lysate Summary

Description

PERK Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC214993

Accession#: NM_004836

Protein Tag: C-MYC/DDK

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
EIF2AK3

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot.

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
122.1 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 PERK Overexpression Lysate

  • DKFZp781H1925
  • EC 2.7.11.1
  • EIF2AK3
  • eukaryotic translation initiation factor 2 alpha kinase 3
  • eukaryotic translation initiation factor 2-alpha kinase 3
  • hsPEK
  • PEK
  • PEKPERKPancreatic eIF2-alpha kinase
  • PERK
  • PRKR-like endoplasmic reticulum kinase
  • WRS

Background

The activation of signal transduction pathways by growth factors, hormones and neurotransmitters is mediated through two closely related MAP kinases, p44 and p42, designated extracellular-signal related kinase 1 (ERK 1) and ERK 2, respectively. ERK proteins are regulated by dual phosphorylation at Tyrosine 204 and 187 and Threonine 177 and 160 residues mapping within a characteristic Thr-Glu-Tyr motif. Phosphorylation at both the Threonine 202 and Tyrosine 204 residues of ERK 1 and Threonine 185 and Tyrosine 187 residues of ERK 2 is required for full enzymatic activation. The structural consequences of dual phosphorylation in ERK 2 include active site closure, alignment of key catalytic residues that interact with ATP, and remodeling of the activation loop. In response to activation, MAP kinases phosphorylate downstream components on serine and threonine. Upstream MAP kinase regulators include MAP kinase kinase (MEK), MEK kinase and Raf-1. The ERK family has three additional members: ERK 3, ERK 5 and ERK 6.

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.

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Video Protocols

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Research Areas for PERK Lysate (NBL1-10177)

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Blogs on PERK.

eIF2alpha - a regulator of global translation in response to cellular stress
Eukaryotic initiation factor 2 (eIF2) regulates global protein translation by binding to Met-tRNA and the 40S ribosome to form the pre-initiation complex. eIF2 is a heterotrimer consisting of alpha, beta, and gamma subunits. The 36kDA eIF2a subuni...  Read full blog post.

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Bioinformatics

Gene Symbol EIF2AK3