Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Predicted:

Bovine (100%), Rat (94%). Backed by our 100% Guarantee.

Applications

Validated:

Western Blot

Cited:

Western Blot, Westen Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all PDK4 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide made to an internal portion of the human PDK4 protein (within residues 100-150). [Swiss-Prot# Q16654]

Reactivity Notes

Immunogen displays the following percentage of sequence identity for non-tested species: rat (94%), bovine (100%)

Localization

Mitochondrion matrix.

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 PDK4 Antibody - BSA Free

Western Blot: PDK4 AntibodyBSA Free [NBP1-07047]

Western Blot: PDK4 AntibodyBSA Free [NBP1-07047]

Western Blot: PDK4 Antibody [NBP1-07047] - Western Blot on human heart protein lysate.
Western Blot: PDK4 AntibodyBSA Free [NBP1-07047]

Western Blot: PDK4 AntibodyBSA Free [NBP1-07047]

Western Blot: PDK4 Antibody [NBP1-07047] - Mouse liver. NuPAGE 4-12%. This image was submitted via customer review.
Western Blot: PDK4 AntibodyBSA Free [NBP1-07047]

Western Blot: PDK4 AntibodyBSA Free [NBP1-07047]

PDK4-Antibody-Western-Blot-NBP1-07047-img0004.jpg
PDK4 Antibody - BSA Free

Western Blot: PDK4 Antibody - BSA Free [NBP1-07047] -

Western Blot: PDK4 Antibody - BSA Free [NBP1-07047] - Genetic manipulations can alter PDH E1 alpha phosphorylation in response to hypoxia.(a) MIA PaCa-2 control (Hg), silenced HIF1 alpha (shHIF1 alpha ) or silenced PDHK1 (shPDHK1) & PDP1 overexpressing cells were incubated for 16 h in normoxia or hypoxia (0.5% O2) & phospho-serine E1 alpha detected by Western blot showing that all three modifications can have an inhibitory effect on hypoxic Ser232 phosphorylation. (b) Increased pSer232-E1 alpha detection after E1 alpha immunoprecipitation from lysates of cells treated as in (a). (c) The same analysis as in (a) after treatment with 16 h 1 mM DMOG. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27498883), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
PDK4 Antibody - BSA Free

Western Blot: PDK4 Antibody - BSA Free [NBP1-07047] -

Western Blot: PDK4 Antibody - BSA Free [NBP1-07047] - Hypoxia inhibits mitochondrial OCR & PDH activity & induces PDHK1 protein & activity.(a) Ratio of oxygen consumption rate (OCR) to extracellular acidification rate (ECAR) measured by Seahorse XF in MIA PaCa-2, PANC-1, & SU.86.86 cell lines in high (25 mM) or low (0.5 mM) glucose incubated overnight with or without 1 mM DMOG. (mean ± SEM, two-tailed Student’s t-test, **p < 0.01, ***p < 0.001) (b) Cell-based PDH activity assay in cells incubated 16 h in normoxia, hypoxia (0.5% O2) or 1 mM DMOG. (mean ± SD, one-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001) (c) Western blots of HIF alpha isoforms, pyruvate dehydrogenase kinase isoforms (PDHKs), phosphatase (PDP1 – lower band *), target phosphorylated serine residues on E1 alpha & total E1 alpha after overnight incubation in normoxia or hypoxia (0.5% O2) at 25 or 5 mM glucose as indicated. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27498883), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for PDK4 Antibody - BSA Free

Application
Recommended Usage

Western Blot

2 ug/ml
Application Notes
In Western blot, a band is seen at ~49 kDa. This product is not recommended for ICC, as nonspecific nuclear 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 3.5 using NBP1-07047 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.1% 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

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: PDK4

PDK4 (pyruvate dehydrogenase kinase isoform 4) is expressed ubiquitously in the mitochondrion matrix of tissues with highest expression seen in liver, heart and skeletal muscles. PDK4 is regulated by glucocorticoids, retinoic acid and insulin, and is found elevated with diabetes, fasting and other conditions where fatty acids act as energy source. Under starvation/diabetic conditions, enhanced glucocorticoids levels activate glucocorticoid receptor thereby recruiting p300/CBP coactivator complex to PDK4 gene where it cooperatively interact with FOXO factors and activated RARs to make a stable transcriptional initiation complex responsible for PDK4 activation. Insulin promotes inactivation and translocation of FOXO factors from nucleus to cytoplasm and thereby suppresses glucocorticoid-stimulated PDK4 gene transcription. PDK4 inhibits the mitochondrial pyruvate dehydrogenase complex (PDC) via its E1 alpha subunit's phosphorylation, thus contributing to glucose metabolism regulation. Insulin resistance is associated with PDC dysregulation in skeletal muscles and excessive insulin on the other hand downregulates PDK4 expression. PDK4 exerts conservation of glucose as well as three-carbon compounds such as lactate, alanine, pyruvate etc. which serve as substrates for gluconeogenesis and this conservation helps maintain euglycemia during starvation, however, exacerbates hyperglycemia in diabetes.

Alternate Names

[Pyruvate dehydrogenase [lipoamide]] kinase isozyme 4, mitochondrial, EC 2.7.11, EC 2.7.11.2, FLJ40832, Pyruvate dehydrogenase kinase isoform 4, pyruvate dehydrogenase kinase, isoenzyme 4, pyruvate dehydrogenase kinase, isozyme 4, pyruvate dehydrogenase, lipoamide, kinase isozyme 4, mitochondrial

Gene Symbol

PDK4

Additional PDK4 Products

Product Documents for PDK4 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 PDK4 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 PDK4 Antibody - BSA Free

Customer Reviews for PDK4 Antibody - BSA Free (2)

3.5 out of 5
2 Customer Ratings
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Customer Images

Showing  1 - 2 of 2 reviews Showing All
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  • PDK4 Antibody
    Name: Thao Luu
    Application: Western Blot
    Sample Tested: Liver
    Species: Mouse
    Verified Customer | Posted 04/05/2018
    PDK4 works very well with super strong signal. That was the intensity we got for probing our mouse liver samples.
    We used the NuPAGE gel 4-12% for our Westerns.
    PDK4 Antibody - BSA Free NBP1-07047
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Human cancer cell whole cell lysate
    Species: Human
    Verified Customer | Posted 08/22/2015

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Protocols

View specific protocols for PDK4 Antibody - BSA Free (NBP1-07047):


Procedure Guide for NBP1-07047 - PDK4 Antibody
Western Blot Protocol
1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 30 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% NFDM + 1% BSA in TBS + Tween, 1 hour at 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-PDK4 primary antibody (NBP1-07047) in blocking buffer and incubate 1 hour at room
temperature.
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 manufacturers
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 manufacturers instructions (Pierce 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.

FAQs for PDK4 Antibody - BSA Free

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  • Q: Which PDK4 antibody do you recommend for immunoblot? I'm working with human cell lines.

    A: NBP1-07047 and NBP1-07049 are our best selling PDK4 antibodies that will meet your mentioned criteria i.e. Immunoblot/Western Blot in Human samples. However, all of our products are 100% guaranteed and you may find a complete list of PDK4 antibodies by searching PDK4 on our website and narrowing your results using the filters on the left-hand side of the screen.

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