Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [4]
- Immunohistochemistry [2]
- Flow cytometry [1]
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- Product number
- MA5-32428 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- A-Raf Recombinant Rabbit Monoclonal Antibody (JJ08-68)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- 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
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- JJ08-68
- 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 A-Raf in Hela cell lysate using a A-Raf Monoclonal antibody (Product # MA5-32428) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of A-Raf in Hela cell lysate using a A-Raf Monoclonal antibody (Product # MA5-32428) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Serine/threonine-protein kinase A-Raf was achieved by transfecting HeLa with Serine/threonine-protein kinase A-Raf specific siRNAs (Silencer® select Product # S577, S576). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the Serine/threonine-protein kinase A-Raf knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with A-Raf Recombinant Rabbit Monoclonal Antibody (JJ08-68) (Product # MA5-32428, 1:2000 ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). 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 Serine/threonine-protein kinase A-Raf.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-A-Raf Recombinant Rabbit Monoclonal Antibody (JJ08-68) (Product # MA5-32428) and a 67 kDa band corresponding to Serine/threonine-protein kinase A-Raf was observed across cell lines. Whole cell extracts (40 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), HT-29 (Lane 3), A-431 (Lane 4), LNCaP (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1:2000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of A-Raf in A431 cells using a A-Raf Monoclonal antibody (Product # MA5-32428) as seen in green. 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 A-Raf in NIH/3T3 cells using a A-Raf Monoclonal antibody (Product # MA5-32428) as seen in green. 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 A-Raf in Hela cells using a A-Raf Monoclonal antibody (Product # MA5-32428) as seen in green. 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 Serine/threonine-protein kinase A-Raf was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with A-Raf Recombinant Rabbit Monoclonal Antibody (JJ08-68) (Product # MA5-32428) 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), (1:2000), 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 Cytosolic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at oil immersion(100X) magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of A-Raf of paraffin-embedded Human kidney tissue using a A-Raf Monoclonal antibody (Product #MA5-32428). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of A-Raf of paraffin-embedded Human breast carcinoma tissue using a A-Raf Monoclonal antibody (Product #MA5-32428). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of A-Raf in Hela cells using a A-Raf Monoclonal Antibody (Product # MA5-32428) 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.