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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
- 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).
- 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 (H+L) 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.
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 (H+L) 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.
