Overview

  • Product name
    Anti-PARK7/DJ1 antibody [EP2815Y]
    See all PARK7/DJ1 primary antibodies
  • Description
    Rabbit monoclonal [EP2815Y] to PARK7/DJ1
  • Host species
    Rabbit
  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications
    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues near the N-terminus of human PARK7/DJ1.

  • Positive control
    • WB: Jurkat, HeLa, NIH3T3 or 293T cell lysate. Human fetal brain, HeLa, Mouse brain, Rat brain tissue lysates IHC-P: Human Lung and Brain tissue. ICC/IF: PANC-1 and Jurkat cell lines. FC: HepG2 cells IP: Mouse brain 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab76008 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 1/5000. Predicted molecular weight: 20 kDa.

For unpurified use at 1/10000 - 1/20000.

IP 1/20.
IHC-P 1/1000.

See IHC antigen retrieval protocols.

Perform heat mediated antigen retrieval using 0.01M Sodium Citrate Buffer, pH 6.0 before commencing with IHC staining protocol.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

For unpurified use at 1/250 - 1/500.

Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/100

ICC/IF 1/50 - 1/500.

Target

  • Function
    Protects cells against oxidative stress and cell death. Plays a role in regulating expression or stability of the mitochondrial uncoupling proteins SLC25A14 and SLC25A27 in dopaminergic neurons of the substantia nigra pars compacta and attenuates the oxidative stress induced by calcium entry into the neurons via L-type channels during pacemaking. Eliminates hydrogen peroxide and protects cells against hydrogen peroxide-induced cell death. May act as an atypical peroxiredoxin-like peroxidase that scavenges hydrogen peroxide. Following removal of a C-terminal peptide, displays protease activity and enhanced cytoprotective action against oxidative stress-induced apoptosis. Stabilizes NFE2L2 by preventing its association with KEAP1 and its subsequent ubiquitination. Binds to OTUD7B and inhibits its deubiquitinating activity. Enhances RELA nuclear translocation. Binds to a number of mRNAs containing multiple copies of GG or CC motifs and partially inhibits their translation but dissociates following oxidative stress. Required for correct mitochondrial morphology and function and for autophagy of dysfunctional mitochondria. Regulates astrocyte inflammatory responses. Acts as a positive regulator of androgen receptor-dependent transcription. Prevents aggregation of SNCA. Plays a role in fertilization. Has no proteolytic activity. Has cell-growth promoting activity and transforming activity. May function as a redox-sensitive chaperone.
  • Tissue specificity
    Highly expressed in pancreas, kidney, skeletal muscle, liver, testis and heart. Detected at slightly lower levels in placenta and brain. Detected in astrocytes, Sertoli cells, spermatogonia, spermatids and spermatozoa.
  • Involvement in disease
    Defects in PARK7 are the cause of Parkinson disease type 7 (PARK7) [MIM:606324]. A neurodegenerative disorder characterized by resting tremor, postural tremor, bradykinesia, muscular rigidity, anxiety and psychotic episodes. PARK7 has onset before 40 years, slow progression and initial good response to levodopa. Some patients may show traits reminiscent of amyotrophic lateral sclerosis-parkinsonism/dementia complex (Guam disease).
  • Sequence similarities
    Belongs to the peptidase C56 family.
  • Post-translational
    modifications
    Sumoylated on Lys-130 by PIAS2 or PIAS4; which is enhanced after ultraviolet irradiation and essential for cell-growth promoting activity and transforming activity.
    Cys-106 is easily oxidized to sulfinic acid.
    Undergoes cleavage of a C-terminal peptide and subsequent activation of protease activity in response to oxidative stress.
  • Cellular localization
    Cytoplasm. Nucleus. Mitochondrion. Under normal conditions, located predominantly in the cytoplasm and, to a lesser extent, in the nucleus and mitochondrion. Translocates to the mitochondrion and subsequently to the nucleus in response to oxidative stress and exerts an increased cytoprotective effect against oxidative damage. Detected in tau inclusions in brains from neurodegenerative disease patients.
  • Information by UniProt
  • Database links
  • Alternative names
    • CAP1 antibody
    • DJ-1 antibody
    • DJ1 antibody
    • DJ1 protein antibody
    • Epididymis secretory sperm binding protein Li 67p antibody
    • FLJ27376 antibody
    • FLJ34360 antibody
    • FLJ92274 antibody
    • HEL S 67p antibody
    • Oncogene DJ1 antibody
    • OTTHUMP00000001348 antibody
    • OTTHUMP00000001349 antibody
    • OTTHUMP00000001350 antibody
    • OTTHUMP00000001351 antibody
    • PARK7 antibody
    • PARK7_HUMAN antibody
    • Parkinson disease (autosomal recessive, early onset) 7 antibody
    • Parkinson disease protein 7 antibody
    • Parkinson protein 7 antibody
    • Protein DJ-1 antibody
    • SP22 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: PARK7/DJ1 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: Human brain tissue lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab76008 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab76008 was shown to specifically react with PARK/DJ1 in wild-type HAP1 cells. No band was observed when PARK/DJ1 knockout samples were used. Wild-type and PARK/DJ1 knockout samples were subjected to SDS-PAGE. ab76008 and ab8245 (loading control to GAPDH) were both diluted 1/10,000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.

  • ab76008 (purified) at 1:20 dilution (0.5µg) immunoprecipitating PARK7/DJ1 in Mouse brain lysate.
    Lane 1 (input): Mouse brain lysate 10µg
    Lane 2 (+): ab76008 & Mouse brain lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76008 in Mouse brain lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling PARK7/DJ1 with Purified ab76008 at 1:1000 dilution (0.11 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling PARK7/DJ1 with Purified ab76008 at 1:500 dilution (0.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling PARK7/DJ1 with Purified ab76008 at 1:20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • All lanes : Anti-PARK7/DJ1 antibody [EP2815Y] (ab76008) at 1/5000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse brain lysates
    Lane 3 : Rat brain lysates

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 20 kDa
    Observed band size: 23 kDa
    why is the actual band size different from the predicted?

  • Anti-PARK7/DJ1 antibody [EP2815Y] (ab76008) at 1/5000 dilution (Purified) + Human fetal brain lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 20 kDa
    Observed band size: 23 kDa why is the actual band size different from the predicted?

  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 (human breast carcinoma) labelling PARK7/DJ1 with purified ab76008 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    Control: PBS only

    This image was generated using the unpurified version of the product. 

  • Overlay histogram showing HepG2 cells stained with ab76008 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76008, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    This image was generated using the unpurified version of the product. 

  • All lanes : Anti-PARK7/DJ1 antibody [EP2815Y] (ab76008) at 1/20000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : NIH3T3 cell lysate
    Lane 4 : 293T cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/1000 dilution

    Predicted band size: 20 kDa
    Observed band size: 23 kDa why is the actual band size different from the predicted?



    This image was generated using the unpurified version of the product. 

  • ab76008, at 1/250 dilution, staining PARK7/DJ1 in human brain by immunohistochemistry using paraffin-embedded tissue.

    This image was generated using the unpurified version of the product. 

References

This product has been referenced in:
  • Wang D  et al. TRPC1 Deletion Causes Striatal Neuronal Cell Apoptosis and Proteomic Alterations in Mice. Front Aging Neurosci 10:72 (2018). Read more (PubMed: 29615894) »
  • Liu Y  et al. Cistanche deserticola polysaccharides protects PC12 cells against OGD/RP-induced injury. Biomed Pharmacother 99:671-680 (2018). WB . Read more (PubMed: 29710464) »
See all 16 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

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Application
Western blot
Sample
Zebrafish Tissue lysate - whole (Brain)
Loading amount
20 µg
Specification
Brain
Treatment
RIPA buffer, 4x Laemmli buffer
Gel Running Conditions
Reduced Denaturing (12% PAGE)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

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Submitted Oct 27 2010

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