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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
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- Product number
- 703699 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- VGLL4 Recombinant Rabbit Monoclonal Antibody (16H37L22)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with Mouse, Rat, Bovine.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 16H37L22
- Vial size
- 100 µg
- Concentration
- 0.5 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
- Knockdown of VGLL4 was achieved by transfecting HCT 116 cells with VGLL4 specific siRNA (Silencer® select Product # s229764 & s229766). Western blot analysis (Fig a) was performed using modified whole cell extracts (1% SDS) from VGLL4 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-VGLL4 Recombinant Rabbit Monoclonal Antibody (Product # 703699, 1:2500 dilution) and Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:5000 dilution). Densitometric analysis of this western blot is shown in the histogram (Fig b). Loss of signal upon siRNA mediated knockdown confirms that the antibody is specific to VGLL4.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-VGLL4 Recombinant Rabbit Monoclonal Antibody (Product # 703699) and a 34 kDa band corresponding to VGLL4 was observed across the cell line tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of HCT 116 (Lane 1), Caco-2 (Lane 2), U87-MG (Lane3), SH-SY5Y (Lane4) and HeLa (Lane 5) were electrophoresed using Novex® NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 (1:2500 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:5000 dilution). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, HCT 116 cells were fixed and permeabilized for detection of endogenous VGLL4 using Anti-VGLL4 Recombinant Rabbit Monoclonal Antibody (Product # 703699, 1:100 dilution) and labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of VGLL4 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, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of VGLL4. Panel e) represents control cells with no primary antibody to assess the background. The images were captured at 60X magnification.
