PRODUCT CODE: ET1704-66

PDK1 Recombinant Rabbit Monoclonal Antibody [JA67-30] (ET1704-66)

  • Recombinant

Applications

  • WB

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of PDK1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1704-66, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: rat heart tissue lysate<br />
Lane 2: mouse heart tissue lysate
  • Western blot analysis of PDK1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1704-66, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: rat heart tissue lysate<br />
Lane 2: mouse heart tissue lysate
  • ICC staining of PDK1 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of PDK1 in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1704-66, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PDK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-66, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-PDK1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-66, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of PDK1 was done on NIH/3T3 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1704-66, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of PDK1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1704-66, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: rat heart tissue lysate
Lane 2: mouse heart tissue lysate

Applications

  • WB

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

PDK1 Recombinant Rabbit Monoclonal Antibody [JA67-30] (ET1704-66)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

Rat heart tissue lysate, mouse heart tissue lysate, Hela, NIH/3T3, human kidney tissue, mouse heart tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JA67-30

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

49 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:2,000

  • ICC:1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

PDK1

SYNONYMS

[Pyruvate dehydrogenase [lipoamide]] kinase isozyme 1, mitochondrial antibody; HGNC:8809 antibody; Mitochondrial pyruvate dehydrogenase kinase isoenzyme 1 antibody; PDH kinase 1 antibody; Pdk1 antibody; PDK1_HUMAN antibody; Pyruvate dehydrogenase kinase isoform 1 antibody; Pyruvate dehydrogenase kinase, isoenzyme 1 antibody

SEQUENCE SIMILARITIES

Belongs to the PDK/BCKDK protein kinase family.

TISSUE SPECIFICITY

Expressed predominantly in the heart. Detected at lower levels in liver, skeletal muscle and pancreas.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated by constitutively activated ABL1, FGFR1, FLT3 and JAK2 (in vitro), and this may also occur in cancer cells that express constitutively activated ABL1, FGFR1, FLT3 and JAK2. Phosphorylation at Tyr-243 and Tyr-244 strongly increases kinase activity, while phosphorylation at Tyr-136 has a lesser effect.

SUBCELLULAR LOCATION

Mitochondrion matrix.

FUNCTION

Mitochondrial pyruvate dehydrogenase (PDH) catalyzes the oxidative decarboxylation of pyruvate and plays a central role in the regulation of homeostasis of carbohydrate fuels in mammals. PDH activity is controlled by a phosphorylation/dephosphorylation cycle, phosphorylation leading to inactivation and dephosphorylation leading to reactivation of PDH. The phosphorylation of PDH is catalyzed by pyruvate dehydrogenase kinase (PDK), the activity of which is stimulated by the products of PDH catalysis. PDK1 consists of alpha and beta subunits; the kinase activity resides in the alpha subunit. Three PDK isoenzymes have been identified in humans (PDK1, 2 and 3) and two have been identified in rodent (PDK1 and 2).