Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [4]
- Flow cytometry [1]
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- Product number
- MA5-32606 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- VAMP2 Recombinant Rabbit Monoclonal Antibody (JM11-00)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- 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.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- JM11-00
- 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 VAMP2 in different lysates using a Monoclonal antibody (Product #MA5-32606) at a dilution of 1:1,000. Positive control: Lane 1: SH-SY5Y, Lane 2: Jurkat.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of VAMP2 in different lysates using a Monoclonal antibody (Product #MA5-32606) at a dilution of 1:1,000. Positive control: Lane 1: SH-SY5Y, Lane 2: Jurkat.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of VAMP2 was achieved by transfecting U-87 MG with VAMP2 specific siRNAs (Silencer® select Product # s13667, s13668). Western Blot analysis (Fig. a) was performed using whole cell extracts (40 µg lysate) from the VAMP2 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with VAMP2 Recombinant Rabbit Monoclonal Antibody (JM11-00) (Product # MA5-32606, 1:1500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10000 dilution). Densitometric analysis of this western Blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Vesicle-associated membrane protein 2. Knockdown is also observed in the ~15 kDa band.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-VAMP2 Recombinant Rabbit Monoclonal Antibody (JM11-00) (Product # MA5-32606) and a 13 kDa band corresponding to Vesicle-associated membrane protein 2 was observed across cell lines tested and is low expressing in Hep G2 and Caco-2 in comparison to U-87 MG and IMR-32. Whole cell extracts (40 µg lysate) of U-87 MG (Lane 1), Hep G2 (Lane 2), T98G (Lane 3), Caco-2 (Lane 4), IMR-32 (Lane 5) and HeLa (Lane 6) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein 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 (1:1500 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). An ~15 kDa band was also detected across the cell lines tested and could correspond to VAMP1 as both VAMPs show high sequence homology.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of VAMP2 in N2A cells using a VAMP2 Monoclonal antibody (Product # MA5-32606) as seen in red. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of VAMP2 in SH-SY5Y cells using a VAMP2 Monoclonal antibody (Product # MA5-32606) as seen in red. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of VAMP2 in SHG-44 cells using a VAMP2 Monoclonal antibody (Product # MA5-32606) as seen in red. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of VAMP2 (Vesicle-associated membrane protein 2) was performed using 70% confluent log phase U-87 MG and Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with VAMP2 Recombinant Rabbit Monoclonal Antibody (JM11-00) (Product # MA5-32606) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2500), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membrane localization of VAMP2 in U-87 MG and no staining in Hep G2 cells (Panel e) which is reported negative for the protein. Panel f represents control U-87 MG cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of VAMP2 in SH-SY5Y cells using a VAMP2 Monoclonal Antibody (Product # MA5-32606) at a dilution of 1:50, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.