Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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- Product number
- MA5-32318 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Cytokeratin 6 Recombinant Rabbit Monoclonal Antibody (SN71-07)
- 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
- SN71-07
- 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 Cytokeratin 6 in human skin lysate using a Cytokeratin 6 Monoclonal antibody (Product # MA5-32318) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Cytokeratin 6 was achieved by transfecting A-431 with Cytokeratin 6 specific siRNAs (Silencer® select Product # s7960, s7950). Western blot analysis (Fig. a) was performed using whole cell extracts from the Cytokeratin 6 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Cytokeratin 6 Recombinant Rabbit Monoclonal Antibody (SN71-07) (Product # MA5-32318, 1:1000 ) 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 Cytokeratin 6. Lysates from A-431 cells transfected with Cytokeratin 5 specific siRNAs (Silencer® select Product # s7956, s7957) (Fig. a, Lanes 4-6) were run as control to ascertain specificity. Densitometric analysis of this western blot is shown in histogram (Fig. c), which shows no decrease in signal.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Cytokeratin 6 Recombinant Rabbit Monoclonal Antibody (SN71-07) (Product # MA5-32318) and a 58kDa band corresponding to Cytokeratin 6 was observed in A431 and HaCaT but not in MCF-7 or NIH:OVCAR-3. Whole cell extracts (30 µg lysate) of A-431 (Lane 1), HaCaT (Lane 2), MCF7 (Lane 3), NIH:OVCAR-3 (Lane 4) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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 (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of Cytokeratin 6 in A431 cells using a Cytokeratin 6 Monoclonal antibody (Product # MA5-32318) 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 Cytokeratin 6 was performed using 70% confluent log phase A-431 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 Cytokeratin 6 Recombinant Rabbit Monoclonal Antibody (SN71-07) (Product # MA5-32318) at 1:200 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 cytoskeletal localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 63X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Cytokeratin 6 of paraffin-embedded Human tonsil tissue using a Cytokeratin-6 Monoclonal antibody (Product #MA5-32318). Counter stained with hematoxylin.