Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody - BSA Free
Novus Biologicals | Catalog # NB100-2383
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human
Predicted:
Chicken (93%), Mouse (100%), Rat (100%). Backed by our 100% Guarantee.
Applications
Validated:
Western Blot, Immunocytochemistry/ Immunofluorescence
Cited:
Western Blot, Immunocytochemistry/ Immunofluorescence
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Format
BSA Free
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Product Specifications
Immunogen
A synthetic peptide made to a C-terminal portion of the human PDK1 protein sequence (between residues 350-436). [Swiss-Prot Q15118]
Reactivity Notes
Predicted to react with mouse and rat based on 100% sequence homology. Immunogen displays the following percentage of sequence identity for non-tested species: chicken (93%).
Localization
Mitochondrion
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Theoretical MW
49 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Scientific Data Images for Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody - BSA Free
Western Blot: Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody [NB100-2383]
Western Blot: PDK-1 Antibody [NB100-2383] - Analysis of HeLa lysates using NB100-2383. Image courtesy of Gregg Semenza - publication number 21620138 (PMID).Immunocytochemistry/ Immunofluorescence: Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody [NB100-2383]
Immunocytochemistry/Immunofluorescence: PDK-1 Antibody [NB100-2383] - Antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).Western Blot: Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody [NB100-2383]
Western Blot: PDK-1 Antibody [NB100-2383] - Detection of PDK-1 in human heart lysate using NB 100-2383 at 2 ug/ml.Applications for Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody - BSA Free
Application
Recommended Usage
Immunocytochemistry/ Immunofluorescence
1:1000-1:2000
Western Blot
2 ug/ml
Application Notes
By Western blot a band is seen at ~49 kDa, representing PDK1 protein. There are also two higher and one lower non-specific molecular weight bands. In ICC/IF, mitochondiral staining was observed in HeLa cells. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Reviewed Applications
Read 2 reviews rated 4.5 using NB100-2383 in the following applications:
Formulation, Preparation, and Storage
Purification
Immunogen affinity purified
Formulation
Tris-Glycine and 0.15M NaCl
Format
BSA Free
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Background: Pyruvate Dehydrogenase Kinase 1/PDK1
PDK1 antibodies are useful tools for glucose metabolism research and mitochondrial metabolism studies.
Alternate Names
EC 2.7.11, EC 2.7.11.2, mitochondrial pyruvate dehydrogenase, lipoamide, kinase isoenzyme 1, PDH kinase 1, PDK1, PDK-1, PKB kinase, Pyruvate dehydrogenase kinase isoform 1, pyruvate dehydrogenase kinase, isoenzyme 1, pyruvate dehydrogenase kinase, isozyme 1, Pyruvate dehydrogenase lipoamide kinase isozyme 1, mitochondrial
Gene Symbol
PDK1
Additional Pyruvate Dehydrogenase Kinase 1/PDK1 Products
Product Documents for Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody - BSA Free
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Product Specific Notices for Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody - BSA Free
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
Citations for Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody - BSA Free
Customer Reviews for Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody - BSA Free (2)
4.5 out of 5
2 Customer Ratings
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Application: Western BlotSample Tested: Human cancer cell whole cell lysateSpecies: HumanVerified Customer | Posted 08/22/2015
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Application: Western BlotSample Tested: HeLa cell lysates, Sample Amount: 50Species: HumanVerified Customer | Posted 01/26/2010
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Protocols
View specific protocols for Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody - BSA Free (NB100-2383):
Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody:
Western Blot Protocol
1. Perform SDS-PAGE (4-12%) on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
3. Rinse membrane with dH2O and then stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% non-fat dry milk + 1% BSA in TBS for 1 hour at room temperature (RT).
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-PDK1 primary antibody (NB 100-2383) in blocking buffer and incubate 1 hour at RT.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions (we used BioFX Super Plus ECL).
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.
Western Blot Protocol
1. Perform SDS-PAGE (4-12%) on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
3. Rinse membrane with dH2O and then stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% non-fat dry milk + 1% BSA in TBS for 1 hour at room temperature (RT).
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
7. Dilute the rabbit anti-PDK1 primary antibody (NB 100-2383) in blocking buffer and incubate 1 hour at RT.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each.
9. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer [TBS + 0.1% Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions (we used BioFX Super Plus ECL).
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided it does not interfere with antibody-antigen binding.
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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