Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody - BSA Free

Novus Biologicals | Catalog # NB100-2383

Novus Biologicals
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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.

Scientific Data Images for Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody - BSA Free

Western Blot: Pyruvate Dehydrogenase Kinase 1/PDK1 Antibody [NB100-2383]

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: 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: 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

Pyruvate dehydrogenase kinase isoform 1 (PDK1) is a component of the mitochondrial multienzyme complex (PHD) which is responsible for carbohydrate fuel regulation. PDK1 inhbits the mitochondrial pyruvate dehydrogenase complex by phosphorylation of the E1 alpha subunit, thus contributing to the regulation of glucose metabolism.
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

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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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Filter By:
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Human cancer cell whole cell lysate
    Species: Human
    Verified Customer | Posted 08/22/2015
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: HeLa cell lysates, Sample Amount: 50
    Species: Human
    Verified 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.

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