Antibody data
- Antibody Data
- Antigen structure
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- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [4]
- Immunohistochemistry [4]
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Validation data
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- Product number
- MA5-32397 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CDK9 Recombinant Rabbit Monoclonal Antibody (SD204-07)
- 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
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- SD204-07
- 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 CDK9 in different lysates using a Monoclonal antibody (Product #MA5-32397) at a dilution of 1:1,000. Positive control: Lane 1: Hela, Lane 2: Jurkat, Lane 3: A431.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Cyclin-dependent kinase 9 was achieved by transfecting HeLa with Cyclin-dependent kinase 9 specific siRNAs (Silencer® select Product # S2835, S2834). Western Blot analysis (Fig. a) was performed using Nuclear enriched extracts from the Cyclin-dependent kinase 9 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with CDK9 Recombinant Rabbit Monoclonal Antibody (SD204-07) (Product # MA5-32397, 1:4000 ) 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 Cyclin-dependent kinase 9.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-CDK9 Recombinant Rabbit Monoclonal Antibody (SD204-07) (Product # MA5-32397) and a 43 and 54 kDa band corresponding to Cyclin-dependent kinase 9 was observed across cell lines tested . Nuclear enriched extracts (50 µg µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), A-431 (Lane 3), Hep G2 (Lane 4), HEK-293 (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 (1:1000) 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 CDK9 in Hela cells using a CDK9 Monoclonal antibody (Product # MA5-32397) 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 CDK9 in A549 cells using a CDK9 Monoclonal antibody (Product # MA5-32397) 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 CDK9 in SW480 cells using a CDK9 Monoclonal antibody (Product # MA5-32397) 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 Cyclin-dependent kinase 9 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 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with CDK9 Recombinant Rabbit Monoclonal Antibody (SD204-07) (Product # MA5-32397) at 1:100 dilution 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 Nuclear localization. Panel e represents control 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
- Immunohistochemical analysis of CDK9 of paraffin-embedded Human lung cancer tissue using a CDK9 Monoclonal antibody (Product #MA5-32397). Counter stained with hematoxylin.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of CDK9 of paraffin-embedded Human breast carcinoma tissue using a CDK9 Monoclonal antibody (Product #MA5-32397). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of CDK9 of paraffin-embedded Human tonsil tissue using a CDK9 Monoclonal antibody (Product #MA5-32397). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of CDK9 of paraffin-embedded Human colon cancer tissue using a CDK9 Monoclonal antibody (Product #MA5-32397). Counter stained with hematoxylin.