DJ-1 antibody | AbD03055
Human anti Human DJ-1 antibody, clone AbD03055 (HCA024) 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.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Human anti Human DJ-1 antibody, clone AbD03055 (HCA024) 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.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Human anti Human DJ-1 (Oxidized at C106)
- Product Type
- Monoclonal Antibody
- Clone
- AbD03055
- Isotype
- HuCAL Fab bivalent
- Specificity
- DJ-1
- Region
- (OXIDIZED AT C106)
Antibody Quality and Performance Guarantee
| Human anti Human DJ-1 antibody, clone AbD03055 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). |
- Target Species
- Human
- Product Form
- A bivalent human recombinant Fab (lambda light chain) 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.
- Preparation
- Metal chelate affinity chromatography
- Source
- E.coli
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.09% Sodium Azide (NaN3)
- Immunogen
- DJ-1 peptide oxidized at C106 (sequence LIAAIC(SO3)AGPTA).
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
- Acknowledgements
- This product and/or its use is covered by claims of U.S. patents, and/or pending U.S. and non-U.S. patent applications owned by or under license to Bio-Rad Laboratories, Inc. See bio-rad.com/en-us/trademarks for details.
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).
His-tag is a registered trademark of EMD Biosciences. - Licensed Use
- For in vitro. research purposes only, unless otherwise specified in writing by Bio-Rad.
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
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 Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |  |
||
| Western Blotting 1 | |
- 1Detection via the Histidine Tag is recommended for Western Blotting applications. Bio-Rad suggest product MCA5995P for this purpose
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.
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | E WB | 125 µl |
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| List Price | Your Price | ||||||
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| Description | Mouse anti Penta Histidine Tag:HRP | ||||||
Source Reference
-
Ooe, H. et al. (2006) Establishment of specific antibodies that recognize C106-oxidized DJ-1.
Neurosci Lett. 404 (1-2): 166-9.
References for DJ-1 antibody
-
Andres-Mateos, E. et al. (2007) DJ-1 gene deletion reveals that DJ-1 is an atypical peroxiredoxin-like peroxidase.
Proc Natl Acad Sci U S A. 104 (37): 14807-12. -
Zhang C et al. (2008) Role of NonO-histone interaction in TNFalpha-suppressed prolyl-4-hydroxylase alpha1.
Biochim Biophys Acta. 1783 (8): 1517-28. -
Bitar, M.S. et al. (2012) Decline in DJ-1 and decreased nuclear translocation of Nrf2 in Fuchs endothelial corneal dystrophy.
Invest Ophthalmol Vis Sci. 53 (9): 5806-13. -
Shadrach KG et al. (2013) DJ-1-dependent regulation of oxidative stress in the retinal pigment epithelium (RPE).
PLoS One. 8 (7): e67983. -
Kim, J. et al. (2015) DJ-1 protects against undernutrition-induced atrophy through inhibition of the MAPK-ubiquitin ligase pathway in myoblasts.
Life Sci. 143: 50-7.
- Synonyms
- PARK7
- RRID
- AB_2160114
- UniProt
- Q99497
- Entrez Gene
- PARK7
- GO Terms
- GO:0005515 protein binding
- GO:0005634 nucleus
- GO:0008219 cell death
- GO:0032091 negative regulation of protein binding
- GO:0060765 regulation of androgen receptor signaling pathway
HCA024
153290
1610
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