703373
antibody from Invitrogen Antibodies
Targeting: CLU
APOJ, CLI, CLU1, CLU2, KUB1, SGP-2, SP-40, TRPM-2
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
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Validation data
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- Product number
- 703373 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Apolipoprotein J Recombinant Rabbit Monoclonal Antibody (12H12L20)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with Mouse, Rat, Bovine.
- Antibody clone number
- 12H12L20
- Concentration
- 0.5 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Apolipoprotein J Recombinant Rabbit Monoclonal Antibody (Product # 703373) and a ~52 kDa band corresponding to Apolipoprotein J was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1), SW-480 (Lane 2), Caco-2 (Lane 3), LNCaP (Lane 4) and PC-3 (Lane 5) were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036) at a 1:4000 dilution. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Apolipoprotein J was achieved by transfecting HeLa cells with Apolipoprotein J specific siRNA (Silencer® select Product # s3158 and Product # s3156). Western blot analysis (Fig a) was performed using whole cell extracts from the Apolipoprotein J knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-Apolipoprotein J Recombinant Rabbit Monoclonal Antibody (Product # 703373) at a 1:1000 dilution and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036) (0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in the histogram (Fig b). Loss of signal upon siRNA mediated knockdown confirms that antibody is specific to Apolipoprotein J.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunocytochemistry analysis, HeLa cells were fixed for detection of endogenous Apolipoprotein J using Anti-Apolipoprotein J Recombinant Rabbit Monoclonal Antibody (Product # 703373) at a 1:100 dilution and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a 1:2000 dilution. Panel a) shows representative cells that were stained for detection and localization of Apolipoprotein J protein (green), Panel b) is stained for nuclei (blue) using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415) at a 1:300 dilution. Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of Apolipoprotein J. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.