Invitrogen

PDK1 Recombinant Rabbit Monoclonal Antibody (JA67-30)

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

1 Published Figure

1 Reference

View all (15) PDK1 antibodies

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Datasheet

Protocols

Questions & Answers

Tech Support

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PDK1 Antibody in Immunocytochemistry (ICC/IF) — 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.

PDK1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of PDK1 of paraffin-embedded Human kidney tissue using a PDK1 Monoclonal antibody (Product #MA5-32702). Counter stained with hematoxylin.

PDK1 Antibody in Western Blot (WB) — 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).

PDK1 Antibody in Flow Cytometry (Flow) — 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).

PDK1 Antibody — Altered expression of Pyruvate dehydrogenase kinase isozyme 1 protein upon cell treatment demonstrated antibody specificity using PDK1 Recombinant Rabbit Monoclonal Antibody (Product # MA5-32702). There was a treatment mediated increase in the expression of Pyruvate dehydrogenase kinase isozyme 1 protein in HeLa cells upon cobalt chloride and deferoxamine mesylate treatments. (https://doi.org/10.1016/j.cmet.2006.02.002).

Product Details

MA5-32702

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:2,000

Immunohistochemistry (IHC)

1:50-1:200

Immunohistochemistry (Paraffin) (IHC (P))

View 1 publication 1 publication

Immunocytochemistry (ICC/IF)

1:50-1:200

Flow Cytometry (Flow)

1:50-1:100

Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Not Applicable

Host/Isotype

Rabbit / IgG

Expression System

HEK293 cells

Class

Recombinant Monoclonal

Type

Antibody

Clone

JA67-30

Immunogen

Synthetic peptide within Human PDK1 aa 16-65

View immunogen

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

TBS, pH 7.4, with 40% Glycerol, 0.05% BSA

Contains

0.05% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2809979

Product Specific Information

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.

Target Information

Serine/threonine kinase that plays a key role in regulation of glucose and fatty acid metabolism and homeostasis via phosphorylation of the pyruvate dehydrogenase subunits PDHA1 and PDHA2. This inhibits pyruvate dehydrogenase activity, and thereby regulates metabolite flux through the tricarboxylic acid cycle, down-regulates aerobic respiration and inhibits the formation of acetyl-coenzyme A from pyruvate. Plays an important role in cellular responses to hypoxia and is important for cell proliferation under hypoxia. Protects cells against apoptosis in response to hypoxia and oxidative stress.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

HIF-1α-Dependent Metabolic Reprogramming, Oxidative Stress, and Bioenergetic Dysfunction in SARS-CoV-2-Infected Hamsters.

International journal of molecular sciences

Jana S,Heaven MR,Stauft CB,Wang TT,Williams MC,D'Agnillo F,Alayash AI

Published figure using PDK1 recombinant monoclonal antibody (Product # MA5-32702) in Immunohistochemistry (Paraffin)

Thu Dec 29 00:00:00 EST 2022

Bioinformatics

Protein Aliases: [Pyruvate dehydrogenase (acetyl-transferring)] kinase isozyme 1, mitochondrial; [Pyruvate dehydrogenase [lipoamide]] kinase isozyme 1, mitochondrial; isoenzyme 1; kinase isoenzyme 1; lipoamide; mitochondrial pyruvate dehydrogenase, lipoamide, kinase isoenzyme 1; PDH kinase 1; PDK p48; pyruvate dehydrogenase; pyruvate dehydrogenase kinase 1; Pyruvate dehydrogenase kinase isoform 1; pyruvate dehydrogenase kinase, isoenzyme 1; pyruvate dehydrogenase kinase, isozyme 1

Gene Aliases: B830012B01; D530020C15Rik; PDHK1; PDK1

UniProt ID: (Human) Q15118, (Mouse) Q8BFP9

Entrez Gene ID: (Human) 5163, (Rat) 116551, (Mouse) 228026

Molecular Function: non-receptor serine/threonine protein kinase