Overview

  • Product name

    Anti-PINK1 antibody [EPR20730]
    See all PINK1 primary antibodies
  • Description

    Rabbit monoclonal [EPR20730] to PINK1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment. This information is considered to be commercially sensitive.

  • Positive control

    • WB: HeLa cells (+/- treatment with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) whole cell lysate; human PINK1 recombinant protein (aa156-507) . ICC/IF: HeLa cells treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours. IP: HeLA cells (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab216144 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Predicted molecular weight: 63 kDa.
ICC/IF 1/500.
IP 1/30.

Target

  • Function

    Protects against mitochondrial dysfunction during cellular stress, potentially by phosphorylating mitochondrial proteins. Involved in the clearance of damaged mitochondria via selective autophagy (mitophagy). It is necessary for PARK2 recruitement to dysfunctional mitochondria to initiate their degradation.
  • Tissue specificity

    Highly expressed in heart, skeletal muscle and testis, and at lower levels in brain, placenta, liver, kidney, pancreas, prostate, ovary and small intestine. Present in the embryonic testis from an early stage of development.
  • Involvement in disease

    Defects in PINK1 are the cause of Parkinson disease type 6 (PARK6) [MIM:605909]. A neurodegenerative disorder characterized by parkinsonian signs such as rigidity, resting tremor and bradykinesia. A subset of patients manifest additional symptoms including hyperreflexia, autonomic instability, dementia and psychiatric disturbances. Symptoms show diurnal fluctuation and can improve after sleep.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications

    Autophosphorylated.
  • Cellular localization

    Mitochondrion outer membrane. Cytoplasm > cytosol.
  • Information by UniProt
  • Database links

  • Alternative names

    • BRPK antibody
    • FLJ27236 antibody
    • mitochondrial antibody
    • PARK 6 antibody
    • PARK6 antibody
    • Phosphatase and Tensin Homolog antibody
    • PINK 1 antibody
    • PINK1 antibody
    • PINK1_HUMAN antibody
    • Protein kinase BRPK antibody
    • PTEN induced putative kinase 1 antibody
    • PTEN induced putative kinase protein 1 antibody
    • PTEN-induced putative kinase protein 1 antibody
    • Serine/threonine kinase PINK1 mitochondrial antibody
    • Serine/threonine protein kinase PINK1 mitochondrial antibody
    • Serine/threonine-protein kinase PINK1 antibody
    see all

Images

  • Anti-PINK1 antibody [EPR20730] (ab216144) at 1/1000 dilution + Human PINK1 recombinant protein (aa156-507), 10 ng

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG H&L (HRP))

    Developed using the ECL technique.

    Predicted band size: 63 kDa
    Observed band size: 38 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Dilution/blocking buffer: 5% NFDM/TBST

  • Immunofluorescent analysis of 4 % paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma)(+/- treatment with 10μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) cells labeling PINK1 with ab216144 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells treated with 10μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    The negative controls are as follows:
    -ve control: PBS, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • All lanes : Anti-PINK1 antibody [EPR20730] (ab216144) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa cells (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Developed using the ECL technique.

    Predicted band size: 63 kDa
    Observed band size: 62 kDa why is the actual band size different from the predicted?


    Exposure time: 5 seconds


    Blocking and dilution buffer: 5% NFDM/TBST

    PINK1 can be induced by CCCP treatment (PMID: 24184327).

  • PINK1 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP. ab141229) for 24 hours) whole cell lysate with ab216144 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216144 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution.

    Lane 1: HeLa (CCCP-treated, ab141229) lysate 10 μg (Input).
    Lane 2: ab216144 IP in HeLa (CCCP-treated, ab141229) lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216144 in HeLa (CCCP-treated, ab141229) whole cell lysate.

    Blocking and dilution buffer: 5% NFDM/TBST.

References

This product has been referenced in:

  • Jia S  et al. Fisetin induces autophagy in pancreatic cancer cells via endoplasmic reticulum stress- and mitochondrial stress-dependent pathways. Cell Death Dis 10:142 (2019). Read more (PubMed: 30760707) »
  • Liu L  et al. Silencing of PINK1 represses cell growth, migration and induces apoptosis of lung cancer cells. Biomed Pharmacother 106:333-341 (2018). Read more (PubMed: 29966978) »
See all 2 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (Hela, ARPE19)
Gel Running Conditions
Reduced Denaturing (4-12% BisTris)
Loading amount
10 µg
Specification
Hela, ARPE19
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Jul 24 2019

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