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  1. Link

    park7dj1-antibody-ep2816y-ab76241.pdf

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Neuroscience Neurology process Neurodegenerative disease Parkinson's disease Parkin / PARK
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Validated using a knockout cell lineRabMAb

Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)

  • Datasheet
Reviews (1) Submit a question References (7)

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Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
  • Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
  • Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
  • Flow Cytometry (Intracellular) - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)

Key features and details

  • Rabbit monoclonal [EP2816Y] to PARK7/DJ1
  • Suitable for: WB, IHC-P, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Knockout
Product image
Human PARK7 (DJ1) knockout HEK-293T cell line (ab266338)
Protein
Product image
Recombinant Human PARK7/DJ1 protein (ab51198)

View more associated products

Overview

  • Product name

    Anti-PARK7/DJ1 antibody [EP2816Y]
    See all PARK7/DJ1 primary antibodies
  • Description

    Rabbit monoclonal [EP2816Y] to PARK7/DJ1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt (Intra)more details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: TF-1, Jurkat, HEK293T, HAP1 and HeLa cell lysates; Human brain nuclear extract tissue lysate; Human brain tissue lysate. Flow Cyt (intra): Jurkat cells. IHC-P: Human brain tissue.
  • 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.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EP2816Y
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Parkinson's disease
    • Parkin / PARK
    • Signal Transduction
    • Signaling Pathway
    • G Protein Signaling
    • Small G Proteins
    • Regulators
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • Nuclear Hormone Receptors
    • Testosterone
    • Signal Transduction
    • Protein Trafficking
    • Chaperones
    • Other Chaperones
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Testosterone
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Chromatin Binding Proteins
    • DNA / RNA binding
    • Cell Biology
    • Other Antibodies
    • Oxidative Stress
    • Cancer
    • Signal transduction
    • G protein signaling
    • Small G proteins
    • Other
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Other
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Neuroscience
    • Diseases

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human PARK7 (DJ1) knockout HEK-293T cell line (ab266338)
  • KO cell lysates

    • Human PARK7 (DJ1) knockout HEK-293T cell lysate (ab257016)
  • KO cell pellets

    • Human PARK7 (DJ1) knockout HEK-293T cell pellet (ab278931)
  • Positive Controls

    • HeLa nuclear extract lysate (ab14655)
    • Jurkat cytoplasmic extract lysate (ab14842)
  • Recombinant Protein

    • Recombinant Human PARK7/DJ1 protein (ab51198)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab76241 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)
1/10000 - 1/20000. Detects a band of approximately 24 kDa (predicted molecular weight: 20 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/50.

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

Notes
WB
1/10000 - 1/20000. Detects a band of approximately 24 kDa (predicted molecular weight: 20 kDa).
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/50.

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

Application notes
Is unsuitable for IP.

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.
  • Target information above from: UniProt accession Q99497 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 11315 Human
    • Entrez Gene: 57320 Mouse
    • Entrez Gene: 117287 Rat
    • Omim: 602533 Human
    • SwissProt: Q99497 Human
    • SwissProt: Q99LX0 Mouse
    • SwissProt: O88767 Rat
    • Unigene: 419640 Human
    • Unigene: 277349 Mouse
    • Unigene: 30105 Rat
    see all
  • 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

  • Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
    Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
    All lanes : Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : PARK7 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : Human brain nuclear fraction tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 20 kDa
    Observed band size: 24 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab76241 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab76241 Anti-PARK7/DJ1 antibody [EP2816Y] was shown to specifically react with PARK7/DJ1 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266338 (knockout cell lysate ab257016) was used. Wild-type and PARK7/DJ1 knockout samples were subjected to SDS-PAGE. ab76241 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
    Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
    All lanes : Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241) at 1/10000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : PARK/DJ1 knockout HAP1 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Human brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 20 kDa
    Observed band size: 24 kDa why is the actual band size different from the predicted?



    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: PARK/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 - ab76241 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.


    ab76241 was shown to specifically react with PARK7/DJ1 in wild-type HAP1 cells. No band was observed when PARK7/DJ1 knockout samples were used. Wild-type and PARK7/DJ1 knockout samples were subjected to SDS-PAGE. ab76241 and ab8245 (loading control to PARK7/DJ1) 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 1hr at room temperature before imaging.

  • Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
    Western blot - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
    All lanes : Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241) at 1/20000 dilution

    Lane 1 : TF-1 cell lysate
    Lane 2 : Jurkat cell lysate
    Lane 3 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 20 kDa
    Observed band size: 24 kDa why is the actual band size different from the predicted?

  • Flow Cytometry (Intracellular) - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
    Flow Cytometry (Intracellular) - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)

    Overlay histogram showing Jurkat cells stained with ab76241 (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 (ab76241, 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 Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARK7/DJ1 antibody [EP2816Y] (ab76241)

    Immunohistochemical analysis of paraffin-embedded human brain tissue using ab76241 at 1/100 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (7)

Publishing research using ab76241? Please let us know so that we can cite the reference in this datasheet.

ab76241 has been referenced in 7 publications.

  • Goljanek-Whysall K  et al. miR-181a regulates p62/SQSTM1, parkin, and protein DJ-1 promoting mitochondrial dynamics in skeletal muscle aging. Aging Cell 19:e13140 (2020). PubMed: 32291905
  • Tsai CY  et al. Redox-active DJ-1 sustains brainstem cardiovascular regulation via maintenance of mitochondrial function during mevinphos intoxication. Neurochem Int 139:104791 (2020). PubMed: 32650030
  • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
  • Di Nottia M  et al. DJ-1 modulates mitochondrial response to oxidative stress: clues from a novel diagnosis of PARK7. Clin Genet N/A:N/A (2016). PubMed: 27460976
  • Prahlad J  et al. Use of cysteine-reactive cross-linkers to probe conformational flexibility of human DJ-1 demonstrates that Glu18 mutations are dimers. J Neurochem 130:839-53 (2014). PubMed: 24832775
  • Besong Agbo D  et al. Development of a capillary isoelectric focusing immunoassay to measure DJ-1 isoforms in biological samples. Anal Biochem 443:197-204 (2013). PubMed: 24055619
  • O'Connell K & Ohlendieck K Proteomic DIGE analysis of the mitochondria-enriched fraction from aged rat skeletal muscle. Proteomics 9:5509-24 (2009). PubMed: 19834913

Customer reviews and Q&As

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Western blot abreview for Anti-PARK7/DJ1 antibody [EP2816Y]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (brain)
Gel Running Conditions
Reduced Denaturing (12,5%)
Loading amount
50 µg
Specification
brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Dr. Polina Vishnyakova

Verified customer

Submitted Mar 29 2019

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