Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [1]
- Flow cytometry [1]
- Other assay [1]
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Validation data
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- Product number
- MA5-29462 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DJ-1 Recombinant Rabbit Monoclonal Antibody (007)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- This product is preservative free. It is recommended to add sodium azide to avoid contamination (final concentration 0.05%-0.1%). Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire. This antibody has specificity for Human PARK7/DJ-1.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 7
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DJ-1 in Lane A: Hela Whole Cell Lysate (30 µg), Lane B: Jurkat Whole Cell Lysate (30 µg) . Samples were probed using a DJ-1 Monoclonal Antibody (Product # MA5-29462) at a 1:1000 dilution, followed by a Goat Anti-Rabbit IgG (H+L), Dylight 800 Secondary Antibody at a 1:10000 dilution. Western blot was performed under reducing conditions. Predicted band size:20 kDa. Observed band size:24 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using DJ-1 Recombinant Rabbit Monoclonal Antibody (7) (Product # MA5-29462) at 1:1,000 dilution. Lane A: Hela Whole Cell Lysate, Lane B: Jurkat Whole Cell Lysate. Lysates/proteins at 30 μg per lane. Secondary Goat Anti-Rabbit IgG H&L (DyLight™ 800) at 1:10,000 dilution. Developed using the Odyssey technique. Performed under reducing conditions. Predicted band size: 20 kDa. Observed band size: 24 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-DJ-1 Monoclonal Antibody, (Product # MA5-29462) and a ~20 kDa band corresponding to DJ-1 was observed across the panel tested. Whole cell extracts (30 µg lysate) of SH-SY5Y treated with Hydrogen peroxide (25mM for 12 hr) (Lane 1), SH-SY5Y (Lane 2), SW480 (Lane 3), U-2-OS (Lane 4), HeLa (Lane 5), Jurkat (Lane 6), MDA-MB-231 (Lane 7), K-562 (Lane 8) and A-431 (Lane 9) were electrophoresed using Novex® NuPAGE® 12% % Bis-Tris gel (Product # NP0341BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-DJ-1 Monoclonal antibody, (Product # MA5-29462) and a ~20 kDa band corresponding to DJ-1 was observed across the Mouse tissues tested except Mouse Placenta and Colon which are reported to be expressing low levels of DJ-1. Tissue extracts (30 µg lysate) of Mouse Skeletal Muscle (Lane1), Mouse Heart (Lane 2), Mouse Kidney (Lane 3), Mouse Placenta (Lane 4) and Mouse Colon (Lane 5) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0341BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DJ-1/PARK7 was performed by loading 20 µg of HeLa wild type (Lane 1), HeLa Cas9 control (Lane 2), HeLa DJ-1/PARK7 knockout (Lane 3) whole cell extracts. The blot was probed with Anti-DJ-1 Recombinant Rabbit Monoclonal Antibody (Product # MA5-29462) (1:1000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036) (1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to DJ-1/PARK7.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence staining of Human DJ-1 in Hela cells. Cells were fixed with 4% PFA, permeabilzed with 0.3% Triton X-100 in PBS, blocked with 10% serum, and incubated with DJ-1 Recombinant Rabbit Monoclonal Antibody (7) (Product # MA5-29462, 1:60) at 37°C 1 hour. Then cells were stained with the Alexa Fluor® 594-conjugated Goat Anti-rabbit IgG secondary antibody (red) and counterstained with DAPI (blue). Positive staining was localized to cytoplasm.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometric analysis of Human DJ-1 expression on HeLa cells. The cells were treated according to manufacturer’s manual, stained with DJ-1 Recombinant Rabbit Monoclonal Antibody (7) (Product # MA5-29462), then a FITC-conjugated Secondary antibody. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DJ-1 Immunoprecipitation using: Lane A: 0.5 mg Jurkat Whole Cell Lysate, Lane B: 0.5 mg Hela Whole Cell Lysate, Lane C: 0.5 mg 293T Whole Cell Lysate 2 µL with DJ-1 Recombinant Rabbit Monoclonal Antibody (7) (Product # MA5-29462) and 15 µL of 50 % Protein G agarose. Primary antibody: DJ-1 Recombinant Rabbit Monoclonal Antibody (7), at 1:1,000 dilution. Secondary antibody: Dylight 800-labeled antibody to rabbit IgG (H+L), at 1:5,000 dilution. Developed using the Odyssey technique. Performed under reducing conditions. Predicted band size: 20 kDa. Observed band size: 20 kDa.