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PDK1 Recombinant Rabbit Monoclonal Antibody [JA67-30]

Usd: 205

Specification

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Catalog# ET1704-66

PDK1 Recombinant Rabbit Monoclonal Antibody [JA67-30]

Application

  • WB

  • IHC-P

  • IF-Tissue

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

PDK1 Recombinant Rabbit Monoclonal Antibody [JA67-30]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human PDK1 aa 16-65 / 436.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Tissue, IP

Molecular Weight

Predicted band size: 49 kDa

Positive Control

HepG2 cell lysate, HEK-293 cell lysate, LNCaP cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse heart tissue lysate, Rat heart tissue lysate, mouse heart tissue, rat heart tissue.

Conjugation

unconjugated

Clone Number

JA67-30

RRID

AB_3070512

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:2,000

  • IF-Tissue

  • 1:200-1:400

  • IP

  • 1-2μg/sample

Target

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).

Background References

1. Du L et al. Overexpression of PIK3CA in murine head and neck epithelium drives tumor invasion and metastasis through PDK1 and enhanced TGF signaling. Oncogene 35:4641-52 (2016).

2. Lelliott CJ et al. Monoclonal antibody targeting of fibroblast growth factor receptor 1c ameliorates obesity and glucose intolerance via central mechanisms. PLoS One 9:e112109 (2014).

Sequence Similarity

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.

UNIPROT

SwissProt: Q15118 Human

SwissProt: Q8BFP9 Mouse

SwissProt: Q63065 Rat

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

Images

  • Western blot analysis of PDK1 on different lysates with Rabbit anti-PDK1 antibody (<a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a>) at 1/5,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (15 µg/Lane)<br />Lane 2: HEK-293 cell lysate (15 µg/Lane)<br />Lane 3: LNCaP cell lysate (15 µg/Lane)<br />Lane 4: NIH/3T3 cell lysate (15 µg/Lane)<br />Lane 5: PC-12 cell lysate (15 µg/Lane)<br />Lane 6: Mouse heart tissue lysate (30 µg/Lane)<br />Lane 7: Rat heart tissue lysate (30 µg/Lane)<br /><br />Predicted band size: 49 kDa<br />Observed band size: 45 kDa<br /><br />Exposure time: 10 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of PDK1 on different lysates with Rabbit anti-PDK1 antibody (ET1704-66) at 1/5,000 dilution.

    Lane 1: HepG2 cell lysate (15 µg/Lane)
    Lane 2: HEK-293 cell lysate (15 µg/Lane)
    Lane 3: LNCaP cell lysate (15 µg/Lane)
    Lane 4: NIH/3T3 cell lysate (15 µg/Lane)
    Lane 5: PC-12 cell lysate (15 µg/Lane)
    Lane 6: Mouse heart tissue lysate (30 µg/Lane)
    Lane 7: Rat heart tissue lysate (30 µg/Lane)

    Predicted band size: 49 kDa
    Observed band size: 45 kDa

    Exposure time: 10 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-66) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of PDK1 on different lysates with Rabbit anti-PDK1 antibody (<a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a>) at 1/1,000 dilution.<br /><br />Lane 1: A549-WT cell lysate<br />Lane 2: A549-KD PDK1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 49 kDa<br />Observed band size: 46 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a>) at 1/1,000 dilution was used in primary antibody diluent at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of PDK1 on different lysates with Rabbit anti-PDK1 antibody (ET1704-66) at 1/1,000 dilution.

    Lane 1: A549-WT cell lysate
    Lane 2: A549-KD PDK1 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 49 kDa
    Observed band size: 46 kDa

    Exposure time: 3 minutes; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-66) at 1/1,000 dilution was used in primary antibody diluent at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-PDK1 antibody (<a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a>) at 1/2,000 dilution for 1 hour 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 with Rabbit anti-PDK1 antibody (ET1704-66) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-66) at 1/2,000 dilution for 1 hour 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 rat heart tissue with Rabbit anti-PDK1 antibody (<a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a>) at 1/2,000 dilution for 1 hour 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 rat heart tissue with Rabbit anti-PDK1 antibody (ET1704-66) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-66) at 1/2,000 dilution for 1 hour 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.

  • PDK1 was immunoprecipitated from 0.2 mg mouse heart tissue lysate with <a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a> at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: Mouse heart tissue lysate (input)<br />Lane 2: <a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a> IP in mouse heart tissue lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1704-66" style="font-weight: bold;text-decoration: underline;">ET1704-66</a> in mouse heart tissue lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 3 minutes; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    PDK1 was immunoprecipitated from 0.2 mg mouse heart tissue lysate with ET1704-66 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1704-66 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: Mouse heart tissue lysate (input)
    Lane 2: ET1704-66 IP in mouse heart tissue lysate
    Lane 3: Rabbit IgG instead of ET1704-66 in mouse heart tissue lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 3 minutes; ECL: K1801

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

Products with the same target and pathway

metafield image

PDK1 Recombinant Rabbit Monoclonal Antibody [JA67-30] - BSA and Azide free

Application: WB,IHC-P,IF-Tissue,IP

Reactivity: Human,Mouse,Rat

Conjugate: unconjugated