Overview

  • Product name

    Anti-PARK7/DJ1 antibody [MJF-R16 (66-5)] - Oxidized
    See all PARK7/DJ1 primary antibodies
  • Description

    Rabbit monoclonal [MJF-R16 (66-5)] to PARK7/DJ1 - Oxidized
  • Host species

    Rabbit
  • Specificity

    This antibody detects the oxidised form of the Park7 protein.
  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human PARK7/DJ1 aa 100-200.
    Database link: Q99497

  • Positive control

    • HeLa whole cell lysate (ab150035), treated with hydrogen peroxide.
  • General notes

    In recent years, a critical need in the Parkinson's Disease (PD) research community has been access to well-characterized antibodies directed against known PD-relevant proteins. The Michael J. Fox Foundation (MJFF) has supported this effort by partnering with Drs. Un Kang and David White (University of Chicago) to help accelerate PD research.
    DJ-1 is widely expressed in the adult mammal and highly conserved between species. Loss-of-function mutations in DJ-1 were recently identified in an autosomal recessive form of Parkinson’s disease (PARK7). Among other roles, DJ-1 protects cells against oxidative stress. Oxidization of the cysteine 106 residue (C106) of DJ-1 occurs as a consequence oxidative stress, but is also necessary to fully activate DJ-1 functions. The oxidation state of DJ-1 C106 appears to lead to distinct roles for DJ-1 in the cellular response to oxidative stress. Oxidation of C106 to the sulfinic (-SO2H) form has been implicated as a necessary step to achieve optimal protective functions of DJ-1, whereas oxidation to the sulfonic form (-SO3H) results in the oxidative destabilization of DJ-1 structure. The sulfonic form has been identified as a major oxidized form in PD brains.

    With the generation of this critical research tool, MJFF hopes to ensure that the role of this modification can be further investigated by all researchers and the relevance of oxidized forms of DJ-1 can be more definitively examined in Parkinson’s disease.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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 ab169520 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 - 1/10000. Predicted molecular weight: 20 kDa.
  • Application notes
    Is unsuitable for ICC/IF or IHC-P.
  • 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

    • All lanes : Anti-PARK7/DJ1 antibody [MJF-R16 (66-5)] - Oxidized (ab169520) at 1/10000 dilution (purified)

      Lane 1 : Untreated HeLa whole cell lysate
      Lane 2 : HeLa whole cell lysate treated with hydrogen peroxide

      Lysates/proteins at 10 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?



      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • All lanes : Anti-PARK7/DJ1 antibody [MJF-R16 (66-5)] - Oxidized (ab169520) at 1/1000 dilution (Unpurified)

      Lane 1 : HeLa cell lysate, untreated
      Lane 2 : HeLa cell lysate, treated with hydrogen peroxide

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 20 kDa



      ab169520 highlights the presence or absence of the oxidized form of this protein; an antibody directed against total PARK7/DJ1 illustrates the presence of PARK7/DJ1 protein in both lanes.

    References

    This product has been referenced in:

    • Gao L  et al. Natrium Benzoate Alleviates Neuronal Apoptosis via the DJ-1-Related Anti-oxidative Stress Pathway Involving Akt Phosphorylation in a Rat Model of Traumatic Spinal Cord Injury. Front Mol Neurosci 12:42 (2019). Read more (PubMed: 30853891) »
    • Jang J  et al. Oxidized DJ-1 Levels in Urine Samples as a Putative Biomarker for Parkinson's Disease. Parkinsons Dis 2018:1241757 (2018). Read more (PubMed: 29887985) »
    See all 4 Publications for this product

    Customer reviews and Q&As

    Answer

    For ab169520, we used a non-denaturing gradient gel without SDS. The specific protocol for cell preparation is given below:

    Hela cells were seeded into 100 mm culture dishes at 5 x 106 cells/dish and grown overnight. To oxidize the cells, growth medium was replaced with Optimem medium containing 0 or 10mM H2O2 and incubated for 1 hour. After incubation, cells were washed 3x with PBS and lysed in a lysis buffer consisting of 1% Triton X-100, 0.25% deoxycholate, 5 mM EDTA, 150 mM NaCl in 50 mM Tris, pH 8.0, supplemented with protease inhibitors pepstatin, leupeptin, and PMSF. Cell lysates were clarified by centrifugation and lysates protein content was determined. Lysates were diluted to 2 ug/uL in load buffer supplemented with 2-mercaptoethanol and heat denatured for 5 min at 100°C.

    While we don't have other antibodies against oxidized DJ1, based on the sequence analysis of ab169520, it has 100% homology with mouse, and the "does not react with mouse" may be due to the unavailability of suitable mouse materials.

    Read More

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