使用敲除细胞株进行验证RabMAb

Anti-PARK7/DJ1抗体[EP2815Y] (ab76008)

概述

性能

应用

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.

应用 Ab评论 说明
WB 1/10000 - 1/20000. Predicted molecular weight: 20 kDa.
IP 1/20.
IHC-P 1/250 - 1/500.

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

Flow Cyt 1/100.

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

ICC/IF 1/50 - 1/500.

靶标

  • 功能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.
  • 组织特异性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.
  • 疾病相关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).
  • 序列相似性Belongs to the peptidase C56 family.
  • 翻译后修饰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.
  • 细胞定位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
  • 数据库链接
  • 别名
    • 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

Anti-PARK7/DJ1 antibody [EP2815Y] 图像

  • 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



  • Predicted band size : 20 kDa

    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 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) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

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

Anti-PARK7/DJ1 antibody [EP2815Y] (ab76008)参考文献

This product has been referenced in:
  • Vázquez-Mayorga E  et al. Novel Redox-Dependent Esterase Activity (EC 3.1.1.2) for DJ-1: Implications for Parkinson's Disease. Int J Mol Sci 17:N/A (2016). Read more (PubMed: 27556455) »
  • Wang B  et al. Essential control of mitochondrial morphology and function by chaperone-mediated autophagy through degradation of PARK7. Autophagy 12:1215-28 (2016). Read more (PubMed: 27171370) »

See all 6 Publications for this product

Product Wall

Thank you for contacting Abcam.
I believe that ab76008 would wwork for you as the peptide for this antibody maps between amino acids ********* of human DJ-1 (UniProt: Q99497).
Please let me know if you have any questions.

Abcam has not validated the combination of species/application used in this Abreview.
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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提交于 Oct 27 2010

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