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Validated using a knockout cell line

Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)

Reviews (3)Q&A (4)References (10)
Immunocytochemistry - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
  • Western blot - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
  • Western blot - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
  • Western blot - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
  • Flow Cytometry (Intracellular) - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)

Key features and details

  • Mouse monoclonal [malphaDJ-1/E2.19] to PARK7/DJ1
  • Suitable for: Flow Cyt (Intra), WB, ICC
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgM

Overview

  • Product name

    Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19]
    See all PARK7/DJ1 primary antibodies

  • Description

    Mouse monoclonal [malphaDJ-1/E2.19] to PARK7/DJ1

  • Host species

    Mouse

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICCmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: ZebrafishDoes not react with: Mouse

  • Immunogen

    Recombinant full length protein corresponding to Human PARK7/DJ1.

  • Positive control

    • WB: HeLa whole cell lysate. Flow Cyt (Intra): HepG2 cells. ICC: HeLa cells.

  • General notes

    This monoclonal antibody to DJ-1 has been knockout validated in Western blot. The expected band for DJ-1 was observed in wild type cells and the band was not seen in knockout cells.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab11251 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
Flow Cyt (Intra)
1/100.

ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.
WB (2)
Use at an assay dependent concentration. Detects a band of approximately 20 kDa.
ICC
1/500.
Notes
Flow Cyt (Intra)
1/100.

ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.
WB
Use at an assay dependent concentration. Detects a band of approximately 20 kDa.
ICC
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

    • Immunocytochemistry - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
      Immunocytochemistry - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)

      ab11251 staining PARK7/DJ1 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab11251 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150121, Goat polyclonal Secondary Antibody to Mouse IgM - mu chain (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

    • Western blot - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
      Western blot - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
      Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251) at 1/500 dilution + HeLa whole cell lysate at 20 µg
    • Western blot - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
      Western blot - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)

      Western blot using clone malphaDJ-1/E2.19 and a beta actin antibody as a loading control.

      The bottom band is PARK7/DJ1, the top band is beta actin.

      Lane 1: 293 cell lysate
      Lane 2: MCF-7 cell lysate
      Lanes 3-7: various different prostate cell lines
      Lane 8: recombinant PARK7/DJ1 (that was used as immunogen for this antibody)

    • Western blot - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
      Western blot - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)

      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 - ab11251  observed at 24 kDa. Red - loading control, ab181602, observed at 37 kDa.

      ab11251 was shown to specifically react with PARK7/DJ1 in wild-type HAP1 cells. No band was observed when knockout samples were used. Wild-type and PARK7/DJ1 knockout samples were subjected to SDS-PAGE. ab11251 and ab181602 (loading control to GAPDH) were diluted at 1/500 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.

    • Flow Cytometry (Intracellular) - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
      Flow Cytometry (Intracellular) - Anti-PARK7/DJ1 antibody [malphaDJ-1/E2.19] (ab11251)
      Overlay histogram showing HepG2 cells stained with ab11251 (red line). The cells were fixed with 80% methanol (5 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 (ab11251, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    References (10)

    Publishing research using ab11251? Please let us know so that we can cite the reference in this datasheet.

    ab11251 has been referenced in 10 publications.

    • Lin Y  et al. High expression of DJ-1 promotes growth and invasion via the PTEN-AKT pathway and predicts a poor prognosis in colorectal cancer. Cancer Med 7:809-819 (2018). WB, IHC-P . PubMed: 29441725
    • Qin LX  et al. BAG5 Interacts with DJ-1 and Inhibits the Neuroprotective Effects of DJ-1 to Combat Mitochondrial Oxidative Damage. Oxid Med Cell Longev 2017:5094934 (2017). PubMed: 28348719
    • Díaz-Casado ME  et al. Melatonin rescues zebrafish embryos from the parkinsonian phenotype restoring the parkin/PINK1/DJ-1/MUL1 network. J Pineal Res 61:96-107 (2016). PubMed: 27064726
    • Principe S  et al. Identification of prostate-enriched proteins by in-depth proteomic analyses of expressed prostatic secretions in urine. J Proteome Res 11:2386-96 (2012). WB . PubMed: 22339264
    • Kim Y  et al. Identification of differentially expressed proteins in direct expressed prostatic secretions of men with organ-confined versus extracapsular prostate cancer. Mol Cell Proteomics 11:1870-84 (2012). WB . PubMed: 22986220
    • Sala GL  et al. The cytotoxic pathway triggered by palytoxin involves a change in the cellular pool of stress response proteins. Chem Res Toxicol 22:2009-16 (2009). WB ; Human . PubMed: 19928802
    • Melle C  et al. Protein profiling of microdissected pancreas carcinoma and identification of HSP27 as a potential serum marker. Clin Chem 53:629-35 (2007). WB, Functional Studies, IHC-Fr ; Human . PubMed: 17303689
    • Bretaud S  et al. p53-dependent neuronal cell death in a DJ-1-deficient zebrafish model of Parkinson's disease. J Neurochem 100:1626-35 (2007). PubMed: 17166173
    • Tang B  et al. Association of PINK1 and DJ-1 confers digenic inheritance of early-onset Parkinson's disease. Hum Mol Genet 15:1816-25 (2006). PubMed: 16632486
    • Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells. J Cell Biochem 92:1221-33 (2004). PubMed: 15258905

    Customer reviews and Q&As

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    1-4 of 4 Q&A

    Question

    Dear Sir/Madam

    I wish to enquire as to the concentration of anti-PARK7/DJ1 antibody (cat # ab11251) (https://www.abcam.com/PARK7DJ1-antibody-malphaDJ-1E219-ab11251.html).

    Regards

    Read More
    Answer

    Thank you for your enquiry.

    I have investigated our records for ab11251. I am sorry toconfirm there is an error on the datsheet and that this antibody is sold as ascites. The datasheet has been updated with the correct information and I apologize for any confusion.

    The quality of our antibodies and datasheets is very important to us, as is any feedback and data we receive. It is regrettable that the information we had on this particular antibody was not of the standard both we and our customers expect from our products.

    With regards to the concentration, unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, and ascites will contain a lot of other proteins, which means the antibody quantification would not be accurate.

    I can confirm that for ascites, concentration of antibody is known to vary between 5 - 10 mg/ml.

    I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you.

    If you have any further questions, please do not hesitate to contact me.

    Read More

    Question

    Previous vial of ab11251 worked in IHC-P, lot 63399 (liquid in vial clear). Vials states Ascites
    New vial of same antibody lot 63399 is not working in IHC-P, liquid in vial is yellowy.Vial does not state ascites.

    Read More
    Answer

    Thank you for contacting us and reporting the problems you have encountered with ab11251. Unfortunately xxx is away today but she has asked me to get in contact with you.

    xxxxhas been investigating how the antibody is produced and purified in house and why there may be a difference in the two vials that you have received and shewill get back to you with this informationas soon as she hasit. In the meantime, if you wouldn'tmind, could you provide a few moredetails of whatproblems you havebeen experiencing with this antibody?I have attached aquestionnaire to this email for this purpose. Thiswill help greatly in investigating this case further and initiating any additional in house testing where necessary. Ifyou could provide images of theantibodyworking andof the latest results that wouldbevery useful.

    As discussed with Kate, although this antibody has not been tested by us for paraffin embedded IHC and would therefore not normally be covered by the Abrpomise, as youhave previously found it to besuccessful in this application we can extend the Abpromise guarantee as a good-will gesture. I am thereforeable to offer you areplacement of thisantibody with adifferent lot,a different antibody altogether (such as https://www.abcam.com/PARK7-DJ1-antibody-ab18257.htmlwhich has been used in paraffin embedded IHC) or if you would prefer, a refund.

    I apologise for the inconvenience this problem has caused and look forward to hearinghow you wouldlike to proceed.

    Read More

    Question

    1 In which dilution the antibody will works best 2 Does the ab11251 fit for immunoprecipitation? thanks for your reply.

    Read More
    Answer

    Thank you for your enquiry. Ab11251 has been tested for application in Western blotting and Immunocytochemistry, and has not yet been tested in any other applications (including IP). For Western blotting, I would recommend starting at a dilution of 1:500, and for ICC I would recommend starting at a dilution of 1:100. Please optimize based upon your results. If you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 Abcam Points will be offered for an image). If you have any additional questions, please contact us again.

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    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 66263 DESCRIPTION OF THE PROBLEM No staining in the brain tissue, staining of the skin SAMPLE zebrafihs whole mount 1-2 days post fertilisation PRIMARY ANTIBODY DJ-1 ab11251 SECONDARY ANTIBODY ABC kit Vector Labs (the dilutions are tested using other antibodies, works nicely) DETECTION METHOD DAB POSITIVE AND NEGATIVE CONTROLS USED omission of the primary and secondary antibodies ANTIBODY STORAGE CONDITIONS reconstituted in water, stored in aliquotes at -20. once thowed, stored at +4. FIXATION OF SAMPLE 4% PFA ANTIGEN RETRIEVAL none PERMEABILIZATION STEP methanol, followed by incubation with PBS+0.3% Triton X100+1%DMSO BLOCKING CONDITIONS same as permeabilization + 4% NGS HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3-4 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES you indicate that this antibody crossreacts with zebrafihs tissues. Could you please specify the conditions and methods used.

    Read More
    Answer

    Thank you very much for your enquiry and for your patience. Ab11251 was originated outside Abcam and according to the originator, they had positive feedback from a researcher who found the antibody to cross-react with zebrafish. That is unfortunately all the information that I have regarding the antibody's cross-reactivity with zebrafish. I saw that you are using ab11251 with PFA fixed brain tissue. To our knowledge, this antibody has only been tested for application in Western blotting and Immunocytochemistry. We don't have any information about how ab11251 works in IHC. At this point I would like to make the following suggestions. The PFA fixative may be modifying the epitope that the antibody recognizes and so I would suggest doing an antigen retrieval step in order to unmask the epitope. Also, you didn't mention which dilutions of ab11251 that you have tried but you may want to increase the concentration. If you have any additional questions, please contact us again.

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