PINK1 Knockout 293T Cell Lysate

Images

 
There are currently no images for PINK1 Lysate (NBP2-65732).

Every product we sell is backed by Novus' 100% Guarantee. If you have used this product, please submit your images and reviews to earn reward points.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB
Concentration
LYOPH

Order Details

PINK1 Knockout 293T Cell Lysate Summary

Preparation
Method
Knockout achieved by using CRISPR/Cas9,-10 bp deletion in exon1
Gene
PINK1

Applications/Dilutions

Application Notes
You will receive 1 vial (100ug) of knockout cell lysate and 1 vial (100ug) of Parental cell lysate. Lysate can be diluted with 1X SDS sample buffer and will be stable at -20 degrees C for 12 months. Minimize freeze-thaw cycles.

Packaging, Storage & Formulations

Storage
Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
Buffer
0.1 mg cell homogenate lyophilized in RIPA buffer made with double-knockout cell lines.
Concentration
LYOPH
Reconstitution Instructions
To use as WB negative control, spin down briefly and resuspend in 100 uL 1xSDS sample buffer (2% SDS, 60 mM Tris-HCl pH 6.8, 10% Glycerol, 0.02% Bromophenol blue, 60 mM beta-mercaptoethanol). Boil the lysate for 3 - 5 minutes before loading it onto gel.

Lysate Details for Array

Type
Knockout 293T Cell
Life Stage
Embryonic
Tissue
Kidney

Notes

Powered by EDIGENE.
Validation of antibody specificity is critical and verification of antibody performance against knockout samples is one way to guarantee that an antibody recognizes a specific target. Novus' KO cell lysate can be used as a negative control for western blots and to confirm the specificity of antibodies.

Alternate Names for PINK1 Knockout 293T Cell Lysate

  • BRPK
  • EC 2.7.11.1
  • FLJ27236
  • PARK6
  • Parkinson disease (autosomal recessive) 6
  • PINK1
  • protein kinase BRPK
  • PTEN induced putative kinase 1
  • PTEN-induced putative kinase protein 1
  • serine/threonine-protein kinase PINK1, mitochondrial

Background

Phosphatase and Tensin Homolog (PTEN) is a tumor suppressor which acts as an antagonist to phosphatidylinositol 3-kinase (PI3K) signaling. PTEN exerts enzymatic activity as a phosphatidylinositol-3,4,5-trisphosphate (PIP3) phosphatase, opposing PI3K activity by reducing availability of PIP3 to proliferating cells. Loss of PTEN function leads to elevated PIP3 and increased activation of PI3K/AKT signaling in many types of cancer.

PINK1 (PTEN induced putative kinase 1) protein contains a N-terminal mitochondrial targeting sequence, putative transmembrane helix, linker region, serine (Ser65)/threonine (Thr257) kinase domain and C-terminal segment. PINK1 is translated in the cytosol, then translocated to the outer mitochondrial membrane where it is rapidly cleaved and degraded as a part of normal mitochondrial function. In damaged (depolarized) mitochondria, PINK1 becomes stabilized and accumulates, resulting in the subsequent phosphorylation of numerous proteins on the mitochondrial surface.

When PINK1 is imported into the cell, mitochondrial processing peptidase, presenilin-associated rhomboid-like protease and AFG3L2 cleave PINK1 and tag it for the ubiquitin-proteasome pathway, keeping low PINK1 protein expression at basal conditions (1,2). Accumulation of PINK1 in mitochondria indicate damage. PINK1 maintains mitochondrial function/integrity, provides protection against mitochondrial dysfunction during cellular stress, and is involved in the clearance of damaged mitochondria via selective autophagy (mitophagy) (3). PINK1 has a theoretical molecular weight of 63 kDa and undergoes proteolytic processing to generate at least two cleaved forms (55 kDa and 42 kDa).

Ultimately PARK2 (E3 Ubiquitin Ligase Parkin) is recruited to the damaged mitochondria where it is activated by 1) PINK-mediated phosphorylation of PARK2 at serine 65, and 2) PARK2 interaction with phosphorylated ubiquitin (also phosphorylated by PINK1 on serine 65) (4,5). There is a strong interplay between Parkin and PINK1, where loss-of-function of human PINK1 results in mitochondrial pathology and can be rescued by Parkin (2,4,5). Mutations in either Parkin or PINK1 alter mitochondrial turnover, resulting in the accumulation of defective mitochondria and, ultimately, neurodegeneration in Parkinson's disease. Mutations in the PINK1 gene located within the PARK6 locus on chromosome 1p35-p36 have been identified in patients with early-onset Parkinson's disease (6).

References

1. Rasool, S., Soya, N., Truong, L., Croteau, N., Lukacs, G. L., & Trempe, J. F. (2018). PINK1 autophosphorylation is required for ubiquitin recognition. EMBO Rep, 19(4). doi:10.15252/embr.201744981

2. Shiba-Fukushima, K., Arano, T., Matsumoto, G., Inoshita, T., Yoshida, S., Ishihama, Y.,... Imai, Y. (2014). Phosphorylation of mitochondrial polyubiquitin by PINK1 promotes Parkin mitochondrial tethering. PLoS Genet, 10(12), e1004861. doi:10.1371/journal.pgen.1004861

3. Vives-Bauza, C., Zhou, C., Huang, Y., Cui, M., de Vries, R. L., Kim, J.,... Przedborski, S. (2010). PINK1-dependent recruitment of Parkin to mitochondria in mitophagy. Proc Natl Acad Sci U S A, 107(1), 378-383. doi:10.1073/pnas.0911187107

4. McWilliams, T. G., Barini, E., Pohjolan-Pirhonen, R., Brooks, S. P., Singh, F., Burel, S.,... Muqit, M. M. K. (2018). Phosphorylation of Parkin at serine 65 is essential for its activation in vivo. Open Biol, 8(11). doi:10.1098/rsob.180108

5. Exner, N., Treske, B., Paquet, D., Holmstrom, K., Schiesling, C., Gispert, S.,... Haass, C. (2007). Loss-of-function of human PINK1 results in mitochondrial pathology and can be rescued by parkin. J Neurosci, 27(45), 12413-12418. doi:10.1523/jneurosci.0719-07.2007

6. Valente, E. M., Bentivoglio, A. R., Dixon, P. H., Ferraris, A., Ialongo, T., Frontali, M.,... Wood, N. W. (2001). Localization of a novel locus for autosomal recessive early-onset parkinsonism, PARK6, on human chromosome 1p35-p36. Am J Hum Genet, 68(4), 895-900. doi:10.1086/319522

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.

Customers Who Viewed This Item Also Viewed...

NB300-270
Species: Hu, Mu, Rt, Ch
Applications: WB, Simple Western, Flow, ICC/IF, IHC-P, IP, In vitro
NBP2-15365
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-Fr, IHC-P
NB300-268
Species: Hu, Mu, Bv, Ce, I, Pl
Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP
AF1438
Species: Hu
Applications: WB, IHC
NB110-41486
Species: Hu, Mu
Applications: WB, Simple Western, IHC
AF1458
Species: Hu, Mu, Rt
Applications: WB, Simple Western, ICC
NB600-1160
Species: Hu, Mu, Rt, Po, Bv, Ze
Applications: WB, Simple Western, ELISA, ICC/IF, IHC, IHC-Fr, IHC-P
AF847
Species: Hu, Mu, Rt
Applications: WB, Simple Western, IHC, CyTOF-ready, ICFlow, KO
NB110-55288
Species: Hu, Mu, Rt, Fi, Pm
Applications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-P, IP
NBP2-21037
Species: Hu
Applications: WB, IHC, IHC-P
NBP2-31361
Species: Hu
Applications: WB, IHC, IHC-P
NBP1-81988
Species: Hu
Applications: IHC, IHC-P
NBP1-84074
Species: Hu
Applications: ICC/IF, IHC, IHC-P
NBP2-75449
Species: Hu, Mu, Rt
Applications: WB, Flow, ICC/IF, IHC, IHC-P, IP
NBP2-02477
Species: Hu, Rt, Ca, Mk
Applications: WB, Flow, ICC/IF, IHC, IHC-P, IP
NBP2-13913
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P
NB100-82245
Species: Hu, Mu, Rt
Applications: WB, Simple Western, IHC, IHC-P
H00002629-M01
Species: Hu, Pl
Applications: WB, ELISA, ICC/IF, IHC-P
NBP2-66383
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P
NBP2-65732
Species: Hu
Applications: WB

Publications for PINK1 Lysate (NBP2-65732) (0)

There are no publications for PINK1 Lysate (NBP2-65732).
By submitting your publication information earn gift cards and discounts for future purchases.

Reviews for PINK1 Lysate (NBP2-65732) (0)

There are no reviews for PINK1 Lysate (NBP2-65732). By submitting a review you will receive an Amazon e-Gift Card or Novus Product Discount.
  • Review with no image -- $10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen
  • Review with an image -- $25/€18/£15/$25 CAD/¥150 Yuan/¥2500 Yen

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol

FAQs for PINK1 Lysate (NBP2-65732) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Other Available Formats

NBP2-65732

Additional PINK1 Products

Bioinformatics Tool for PINK1 Lysate (NBP2-65732)

Discover related pathways, diseases and genes to PINK1 Lysate (NBP2-65732). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
Visit Tool

Diseases for PINK1 Lysate (NBP2-65732)

Discover more about diseases related to PINK1 Lysate (NBP2-65732).
 

Pathways for PINK1 Lysate (NBP2-65732)

View related products by pathway.

PTMs for PINK1 Lysate (NBP2-65732)

Learn more about PTMs related to PINK1 Lysate (NBP2-65732).
 

Research Areas for PINK1 Lysate (NBP2-65732)

Find related products by research area.

Blogs on PINK1. Showing 1-10 of 12 blog posts - Show all blog posts.

Losing memory: Toxicity from mutant APP and amyloid beta explain the hippocampal neuronal damage in Alzheimer's disease
By Jamshed Arslan Pharm.D. Alzheimer's disease (AD) is an irreversible brain disorder that destroys memory and thinking skills. The telltale signs of AD brains are extracellular deposits of amyloid beta (a polypepti...  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.

The identification of dopaminergic neurons using Tyrosine Hydroxylase in Parkinson's research and LRRK2
Tyrosine hydroxylase (TH) is a crucial enzyme involved in the biosynthesis of dopamine, norepinephrine and epinephrine in the brain.  Specifically, TH catalyzes the conversion of l-tyrosine to l-dihydroxyphenylalanine (l-dopa).  The importance of t...  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 and its role in Parkinson's disease
PINK1 (PTEN induced putative kinase 1) is a mitochondrial serine/threonine kinase which maintains mitochondrial function/integrity, provides protection against mitochondrial dysfunction during cellular stress, potentially by phosphorylating mitochondr...  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.

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...  Read full blog post.

Showing 1-10 of 12 blog posts - Show all blog posts.

Customers Who Bought This Also Bought

LRRK2 Antibody
NB300-268
PINK1 Antibody
BC100-494
DRP1 Antibody
NB110-55288

Contact Information

Product PDFs

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Review this Product

Be the first to review our PINK1 Knockout 293T Cell Lysate and receive a gift card or discount.

Bioinformatics

Gene Symbol PINK1