Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [3]
- Immunohistochemistry [2]
- Flow cytometry [1]
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- Product number
- MA5-32702 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PDK1 Recombinant Rabbit Monoclonal Antibody (JA67-30)
- 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
- JA67-30
- 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 PDK1 in rat heart tissue lysate using a PDK1 Monoclonal antibody (Product # MA5-32702) at a dilution of 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Pyruvate dehydrogenase kinase isozyme 1 protein Monoclonal Antibody (Product # MA5-32702) and 49.4 kDa band corresponding to Pyruvate dehydrogenase kinase isozyme 1 protein was observed across cell lines tested. Hypoxia conditions such as cobalt chloride and deferoxamine mesylate treatments are known to increase the expression of Pyruvate dehydrogenase kinase isozyme 1 protein. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with cobalt chloride (150uM for 24 hr) (Lane 2) and HeLa treated with deferoxamine mesylate (10uM for 20 hr) (Lane 3), Hep G2 (Lane 4), Mouse heart (Lane 5) and Mouse liver (Lane 6), Rat heart (Lane 7) and Rat Liver (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0342BOX). 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 dilution) and detected by Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) 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 PDK1 in Hela cells using a PDK1 Monoclonal antibody (Product # MA5-32702) 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 PDK1 in NIH-3T3 cells using a PDK1 Monoclonal antibody (Product # MA5-32702) 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 PDK1 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 1 hour at room temperature. The cells were labeled with PDK1 Recombinant Rabbit Monoclonal Antibody (Product # PA5-32702) at 1:100 in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 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). Panel d represents the merged image showing Cytoplasmic 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 PDK1 of paraffin-embedded Human kidney tissue using a PDK1 Monoclonal antibody (Product #MA5-32702). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of PDK1 of paraffin-embedded Mouse heart tissue using a PDK1 Monoclonal antibody (Product #MA5-32702). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of PDK1 in NIH-3T3 cells using a PDK1 Monoclonal Antibody (Product # MA5-32702) at a dilution of 1:100, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black).