Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
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Validation data
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- Product number
- PA5-19674 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CDK9 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with mouse, rat, chicken and cow based on sequence homology.
- Reactivity
- Human, Mouse, Xenopus
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 200 µL
- Concentration
- 0.4 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 A431 Whole Cell Lysate using Product # PA5-19674, Cdk9 primary antibody at a dilution of 1:250 (lane 1). Staining of HEK293 Whole Cell Lysate at a dilution of 1:250 (lane 2). Blot treated with a secondary IR Dye680-conjugated Goat polyclonal anti-Rabbit antibody was used at a dilution of 1:10000.
- Submitted by
- 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 Polyclonal Antibody (Product # PA5-19674, 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 Cyclin-dependent kinase 9.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-CDK9 Polyclonal Antibody (Product # PA5-19674) and a ~43 kDa band corresponding to Cyclin-dependent kinase 9 was observed across cell lines and tissues tested . Nuclear enriched extracts (50 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), A-431 (Lane 3), Hep G2 (Lane 4), HEK-293 (Lane 5), Mouse Skeletal Muscle (Lane 6), Rat Kidney (Lane 7) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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 (1:500) 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
- Immunofluorescent staining of HeLa cells using Product # PA5-19674, anti-Cdk9 antibody. The cells were fixed with PFA (4%) for 10 minutes, permabilised with BSA (1%), normal goat serum (10%) and glycine (0.3 M) in 0.1% T-BST for 20 minutes and exposed to the primary antibody at a concentration of 1 µg/mL for 1 hour at room temp. The secondary antibody was a 448 fluorescence conjugated Goat anti-rabbit IgG (green) at a dilution of 1:1000. A WGA- 594 fluorescent conjugated stain was used to label plasma membranes (red) and the nuclei stain was DAPI (blue).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent staining of HeLa cells using Product # PA5-19674, anti-Cdk9 antibody. The cells were fixed with PFA (4%) for 10 minutes, permabilised with BSA (1%), normal goat serum (10%) and glycine (0.3 M) in 0.1% T-BST for 20 minutes and exposed to the primary antibody at a concentration of 1 µg/mL for 1 hour at room temp. The secondary antibody was a 448 fluorescence conjugated Goat anti-rabbit IgG (green) at a dilution of 1:1000. A WGA- 594 fluorescent conjugated stain was used to label plasma membranes (red) and the nuclei stain was DAPI (blue).