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Parkin Overexpression Lysate

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Western Blot: Parkin Overexpression Lysate (Adult Normal) [NBL1-14105] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Parkin.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

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Parkin Overexpression Lysate Summary

Description

Parkin Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC216186

Accession#: NM_013988

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
PRKN

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
35.5 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.

Alternate Names for Parkin Overexpression Lysate

  • AR-JP
  • E3 ubiquitin ligase
  • E3 ubiquitin-protein ligase parkin
  • EC 6.3.2.-
  • LPRS2
  • PARK2
  • parkin 2
  • Parkin
  • Parkinson disease protein 2
  • Parkinson juvenile disease protein 2
  • parkinson protein 2, E3 ubiquitin protein ligase (parkin)
  • PDJ
  • PDJjuvenile) 2, parkin
  • PRKN

Background

Parkinson's disease is a common neurodegenerative disease with complex clinical features. Mutations in the gene, Parkin (PARK2), appear to be responsible for the pathogenesis of autosomal recessive juvenile Parkinsonism. Parkin plays a role in the ubiquitin-mediated proteolytic pathway by removal and/or detoxification of abnormally folded or damaged protein. Loss of this ubiquitin ligase activity appears to be the mechanism underlying pathogenesis of Parkin. Parkin may protect neurons against alpha synuclein toxicity, proteasomal dysfunction, gpr37 accumulation, and kainate-induced excitotoxicity. It may play a role in controlling neurotransmitter trafficking at the presynaptic terminal and in calcium-dependent exocytosis. Parkin also regulates cyclin e during neuronal apoptosis and may represent a tumor suppressor gene.

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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Publications for Parkin Lysate (NBL1-14105) (0)

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Product General Protocols

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Video Protocols

WB Video Protocol

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Research Areas for Parkin Lysate (NBL1-14105)

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Blogs on Parkin.

Understanding Mitophagy Mechanisms: Canonical PINK1/Parkin, LC3-Dependent Piecemeal, and LC3-Independent Mitochondrial Derived Vesicles
By Christina Towers, PhD What is Mitophagy?The selective degradation of mitochondria via double membrane autophagosome vesicles is called mitophagy. Damaged mitochondria can generate harmful amounts of reactive ox...  Read full blog post.

New Players in the Mitophagy Game
By Christina Towers, PhD Mitochondrial turn over via the lysosome, otherwise known as mitophagy, involves engulfment of mitochondria into double membrane autophagosomes and subsequent fusion with lysosomes. Much is al...  Read full blog post.

Optogenetic Control of Mitophagy: AMBRA1 based mitophagy switch
By Christina Towers, PhD Mitophagy in the BrainSelective autophagic degradation of damaged mitochondria, known as mitophagy, has been described as a cyto-protective process. Accordingly, defects in mitophagy h...  Read full blog post.

There's an autophagy for that!
By Christina Towers, PhDA critical mechanism that cells use to generate nutrients and fuel metabolism is through a process called autophagy.  This process is complex and involves over 20 different proteins, most of which are highly conserved acro...  Read full blog post.

The role of Parkin and autophagy in retinal pigment epithelial cell (RPE) degradation
The root of Parkinson’s disease (PD) points to a poorly regulated electron transport chain leading to mitochondrial damage, where many proteins need to work cohesively to ensure proper function.  The two key players of this pathway are PINK1, ...  Read full blog post.

Parkin - Role in Mitochondrial Quality Control and Parkinson's Disease
Parkin/PARK2 is a cytosolic enzyme which gets recruited to cellular mitochondria damaged through depolarization, ROS or unfolded proteins accumulation, and exert protective effects by inducing mitophagy (mitochondrial autophagy). Parkin induces mit...  Read full blog post.

PINK1 - performing mitochondrial quality control and protecting against Parkinson’s disease
PTEN-induced putative kinase 1 (PINK1) is a serine/threonine kinase with important functions in mitochondrial quality control. Together with the Parkin protein, PINK1 is able to regulate the selective degradation of damaged mitochondria through aut...  Read full blog post.

PINK1: All work and no fun
The protein PINK1 is a mitochondrial-located serine/threonine kinase (PTK) that maintains organelle function and integrity. It not only protects organelles from cellular stress, but it also uses the selective auto-phagocytosis process for cleaning and...  Read full blog post.

PINK1: Promoting Organelle Stability and Preventing Parkinson's disease
PINK1 is a protein serine/threonine kinase (PTK) that protects the organelles from cellular stress and controls selective autophagy to clear damage. Exner, et al. were among the first to report that PINK1 deficiency in humans was linked to autosomal r...  Read full blog post.

Novus Antibodies Highlighted in Parkinson's Disease Research
Identified almost two centuries ago, Parkinson's disease is a neurodegenerative disorder that afflicts an estimated 4-6 million worldwide (www.parkinsons.org). The prevalence of Parkinson's disease is expected to grow considerably as the average age o...  Read full blog post.

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Bioinformatics

Gene Symbol PRKN