DJ-1 Antibody | 3055

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DJ-1 Antibody | 3055 gallery image 1

Western blot demonstrating specifity of HCA024 for DJ-1 oxidized at C106. 293T cells were transfected with various wild-type and cysteine substitution mutants of DJ-1, and extracts were then immunoprecipitated with anti-DYKDDDDK antibody followed by Western blot with HCA024

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DJ-1 Antibody | 3055 gallery image 2

Detection of oxidized peptide compared to non oxidized peptide in ELISA with Human anti Human DJ-1 oxidized at C106 (HCA024)

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DJ-1 Antibody | 3055 gallery image 3

Published customer image:
Human anti Human DJ-1 antibody, clone 3055 used for the detection of oxidized DJ-1 by immunofluorescence and western blotting
Image caption:
Presence of oxDJ-1 in RPE cells subjected to oxidative stress. ARPE-19 monolayers were treated with increasing concentrations (0 to 800 µM) of H2O2 for 1 hr (A) and 18 hs (B), harvested, and analyzed by immunoblot assay with oxDJ-1 antibody (upper panel). Protein loadings were confirmed in replicate blots probed with GAPDH (lower panel). Each lane contained 20 µg of protein. A dose response is observed when cells are exposed to increasing concentrations of H2O2 for 1 h (A, lanes 1 to 6) and 18 hrs (B, lanes 7 to 12). Confocal immunofluorescence staining of baseline ARPE-19 cultures (C) fixed before extraction with Triton X-100 and labeling with oxDJ-1 antibodies revealed absence of oxDJ-1. However, oxDJ-1 is observed in the cytoplasm (arrows) and perinuclear area (arrowheads) of RPE cells exposure to 400 µM H2O2 for 1 h (D) and 18 hrs (E). Cell nuclei were labeled with TO-PRO-3. Scale bar = 20 µm.

From: Shadrach KG, Rayborn ME, Hollyfield JG, Bonilha VL (2013) DJ-1-Dependent Regulation of Oxidative Stress in the Retinal Pigment Epithelium (RPE).
PLoS ONE 8(7): e67983.

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DJ-1 Antibody | 3055 gallery image 4

Published customer image:
Human anti Human DJ-1 antibody, clone 3055 used for the detection of oxidized DJ-1 by western blotting
Image caption:
Increased levels of DJ-1 and oxDJ-1 in RPE lysates and tissue from AMD donors. Lysates of human RPE isolated from non-AMD (A to C lanes 1 to 5) and AMD donors (A to C, lanes 6 to 10) were harvested and analyzed by immunoblot assay with DJ-1 antibody (A) and oxDJ-1 antibody (B). Each lane contained 20 µg of protein. Protein loads were confirmed in replicate blots probed with GAPDH (C). Immunoblots of lysates revealed that AMD RPE displayed significant increased immunoreactivity of both DJ-1 (A) and oxDJ-1 (B) when compared to non-AMD RPE lysates. Quantification of immunoblots demonstrated a 1.7 and 4 fold increase in the expression levels of DJ-1 and oxDJ-1, respectively (D). Blue columns = non-AMD; Red columns = AMD. Data is expressed as mean relative signal intensity ± SEM (n = 8). Asterisks denote statistical significance compared with non-AMD RPE (*p = p = 0.0098 for DJ-1 and **p = 0.0058 for oxDJ-1). Alternatively, cryosections of different non-AMD (E–H) and AMD (I to L) donors with geographic atrophy, and isolated Bruch’s membrane (BM) and choroid from two different AMD donors were (M to P) labeled with DJ-1 antibody. Negative control sections were reacted with DJ-1 antibody pre-absorbed with lysates of cells overexpressing DJ-1 and shoed no DJ-1 labeling (E, G, I, K, M, O). DJ-1 labeling was detected mostly in the RPE nuclei (arrowheads) but also in the cytoplasm of non-AMD donors (F and H, arrows). Significantly more DJ-1 was detected all over the cytoplasm of RPE cells from two different AMD donors (J and L, arrows), while DJ-1 was diffusely distributed in isolated BM and in drusen (E, G, asterisks). Scale bars (E to L) = 10 µm; (M to P) = 50 µm.

From: Shadrach KG, Rayborn ME, Hollyfield JG, Bonilha VL (2013) DJ-1-Dependent Regulation of Oxidative Stress in the Retinal Pigment Epithelium (RPE).
PLoS ONE 8(7): e67983.

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  • Human anti Human DJ-1 (Oxidized At C106)
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Monoclonal Antibody
  • Clone
    3055
  • Isotype
    HuCAL Fab bivalent
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    HCA024E, WBdatasheet pdfdatasheet pdf0.1 mg
    HCA024
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Human anti Human DJ-1 antibody, clone 3055 recognizes the human DJ-1 oncogene only when it is oxidized on cysteine C106. DJ-1 plays roles in transcriptional regulation and reaction to anti-oxidative stress. The oxidation status of DJ-1, particularly at C106, modulates the function of DJ-1 (Canet-Avilés et al. 2004). De-regulation of DJ-1 oxidation appears to be connected to the onset of conditions such as Parkinson's Disease (Abou-Sleiman et al. 2003).
    • Intended Use
    • Target Species
      Human
    • Product Form
      A liquid, bivalent human recombinant Fab selected from the HuCAL® GOLD phage display library. Expressed in E. coli and purified using NiNTA affinity chromatography. This Fab fragment is dimerized via a helix-turn-helix motif. The antibody is tagged with a myc-tag (EQKLISEEDL) and a his-tag (HHHHHH) at the C-terminus of the antibody heavy chain.
    • Reconstitution
    • Preparation
      Metal chelate affinity chromatography.
    • Preservative Stabilisers
      0.01% Thiomersal
    • Immunogen
      DJ-1 peptide oxidized at C106 (sequence LIAAIC(SO3)AGPTA).
    • Purity
    • Approx. Protein Concentrations
      Antibody concentration 0.5 mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
    • Storage
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      12 months from date of despatch.
    • GO Terms
      protein binding
      cell death
      negative regulation of protein binding
      regulation of androgen receptor signaling pathway
      nucleus
    • UniProt
    • Entrez Gene
    • Acknowledgements
      Sold under license of U.S. Patents 6,300,064, 6,696,248, 6,708,484, 6,753,136, European patent 0,859,841 and corresponding patents.
      "By purchasing this product, you agree that your Name and Affiliation will be sent to Dr. Ariga, Hokkaido University, who first published this antibody in the journal Neuroscience Letters (Vol. 404, pp.166-169, 2006)."
    • Licensed Use
      For in vitro research purposes only, unless otherwise specified in writing by Bio-Rad.
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      ELISA
      Western Blotting

    • Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
    • Technical Advice
      Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use

    Additional DJ-1 Antibody Formats

    Formats Clone Applications Sizes available
    DJ-1 Antibody : Purified 3055 E, WB 0.1 mg
    • Copyright © 2016 Bio-Rad

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      Recommended Negative Isotype Control

        Useful Reagents

          Recommended Positive Controls

            Histology Controls

              • Application NameReference Images
                Western Blotting

              References

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