使用敲除细胞株进行验证

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

概述

  • 产品名称
    Anti-PARK7/DJ1抗体[malphaDJ-1/E2.19]
    参阅全部 PARK7/DJ1 一抗
  • 描述
    小鼠单克隆抗体[malphaDJ-1/E2.19] to PARK7/DJ1
  • 特异性
    This clone has been shown to specifically recognise a fusion protein of PARK7/DJ1. It also recognises a FLAG-tagged PARK7/DJ1 expressed in eukaryotic cells. A single band is seen when Western blotting in optimised conditions with this clone. However, overloading the gel or using low dilutions can cause other bands to appear.
  • 经测试应用
    适用于: Flow Cyt, IHC-Fr, ICC/IF, WB, ICC, IHC-FoFrmore details
  • 种属反应性
    与反应: Human, Zebrafish
    不与反应: Mouse
  • 免疫原

    Recombinant full length protein (Human).

  • 阳性对照
    • WB: HeLa whole cell lysate. Flow Cytometry: HepG2 cells.
  • 常规说明

    This antibody clone is manufactured by Abcam.

    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.

性能

应用

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.

应用 Ab评论 说明
Flow Cyt 1/100. ab91545-Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.
IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/500.
WB Use at an assay dependent concentration. Detects a band of approximately 20 kDa.
ICC Use at an assay dependent concentration.
IHC-FoFr 1/1000.

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

图片

  • Western blot using ab11251 at 1/500 on HeLa whole cell lysate (20µg/lane).

    Western blot using ab11251 at 1/500 on HeLa whole cell lysate (20µg/lane).
  • 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)

  • 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.

  • ICC/IF image of ab11251 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab11251, 1:500 dilution) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • 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.

文献

This product has been referenced in:
  • 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). Read more (PubMed: 28348719) »
  • 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 . Read more (PubMed: 22339264) »

See all 8 Publications for this product

客户评价及客户问答

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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 apol...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (normal fibroblasts)
Loading amount
40 µg
Specification
normal fibroblasts
Gel Running Conditions
Reduced Denaturing (12 % polyacrylamide gel)
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Username

Dr. Paule Benit

Verified customer

提交于 May 03 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (normal human fibroblasts)
Specification
normal human fibroblasts
Fixative
Formaldehyde
Permeabilization
Yes - 1X PBS 0.25% Triton
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 37°C
Username

Dr. Paule Benit

Verified customer

提交于 Apr 23 2010

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 1:5...

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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 a...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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