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Pyruvate Dehydrogenase E2 Recombinant Rabbit Monoclonal Antibody [JE34-38]

Usd: 385

Specification

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Catalog# HA721267

Pyruvate Dehydrogenase E2 Recombinant Rabbit Monoclonal Antibody [JE34-38]

Application

  • WB

  • IHC-P

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

Pyruvate Dehydrogenase E2 Recombinant Rabbit Monoclonal Antibody [JE34-38]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human Pyruvate Dehydrogenase E2 aa 574-624/647.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Molecular Weight

Predicted band size: 69 kDa

Positive Control

HL-60 cell lysate, Mouse heart tissue lysate, Mouse liver tissue lysate, Rat liver tissue lysate, HAP1 cell lysate, human testis tissue, mouse cerebellum tissue, rat brain tissue, rat hippocampus tissue.

Conjugation

unconjugated

Clone Number

JE34-38

RRID

AB_3072384

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

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

  • 1:1,000-1:2,000

  • IHC-P

  • 1:200

Target

Function

Dihydrolipoyl transacetylase (or dihydrolipoamide acetyltransferase) is an enzyme component of the multienzyme pyruvate dehydrogenase complex. The pyruvate dehydrogenase complex is responsible for the pyruvate decarboxylation step that links glycolysis to the citric acid cycle. This involves the transformation of pyruvate from glycolysis into acetyl-CoA which is then used in the citric acid cycle to carry out cellular respiration. There are three different enzyme components in the pyruvate dehydrogenase complex. Pyruvate dehydrogenase (EC 1.2.4.1) is responsible for the oxidation of pyruvate, dihydrolipoyl transacetylase (this enzyme; EC 2.3.1.12) transfers the acetyl group to coenzyme A (CoA), and dihydrolipoyl dehydrogenase (EC 1.8.1.4) regenerates the lipoamide. Because dihydrolipoyl transacetylase is the second of the three enzyme components participating in the reaction mechanism for conversion of pyruvate into acetyl CoA, it is sometimes referred to as E2. In humans, dihydrolipoyl transacetylase enzymatic activity resides in the pyruvate dehydrogenase complex component E2 (PDCE2) that is encoded by the DLAT (dihydrolipoamide S-acetyltransferase) gene.

Background References

1. Chen S et al. The phytochemical hyperforin triggers thermogenesis in adipose tissue via a Dlat-AMPK signaling axis to curb obesity. Cell Metab. 2021 Mar

2. Chen Q et al. PM2.5 promotes NSCLC carcinogenesis through translationally and transcriptionally activating DLAT-mediated glycolysis reprograming. J Exp Clin Cancer Res. 2022 Jul

Subcellular Location

Mitochondrion matrix.

UNIPROT

SwissProt: P10515 Human

SwissProt: Q8BMF4 Mouse

SwissProt: P08461 Rat

Synonyms

70 kDa mitochondrial autoantigen of primary biliary cirrhosis antibody

Dihydrolipoamide acetyltransferase component of pyruvate dehydrogenase complex antibody

Dihydrolipoamide S Acetyltransferase antibody

Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex antibody

dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex mitochondrial antibody

DLAT antibody

DLTA antibody

E2 antibody

E2 component of pyruvate dehydrogenase complex antibody

M2 antigen complex 70 kDa subunit antibody

Expand

Images

  • Western blot analysis of Pyruvate Dehydrogenase E2 on different lysates with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (<a href="/products/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/1,000 dilution.<br /><br />Lane 1: HL-60 cell lysate (20 µg/Lane)<br />Lane 2: Mouse heart tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 69 kDa<br />Observed band size: 60 kDa<br /><br />Exposure time: 4 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/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/1,000 dilution was used in 5% NFDM/TBST 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 Pyruvate Dehydrogenase E2 on different lysates with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (HA721267) at 1/1,000 dilution.

    Lane 1: HL-60 cell lysate (20 µg/Lane)
    Lane 2: Mouse heart tissue lysate (40 µg/Lane)

    Predicted band size: 69 kDa
    Observed band size: 60 kDa

    Exposure time: 4 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 (HA721267) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Pyruvate Dehydrogenase E2 on different lysates with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (<a href="/products/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/2,000 dilution.<br /><br />Lane 1: Mouse liver tissue lysate<br />Lane 2: Rat liver tissue lysate<br /><br />Lysates/proteins at 40 µg/Lane.<br /><br />Predicted band size: 69 kDa<br />Observed band size: 60 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/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/2,000 dilution was used in 5% NFDM/TBST 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 Pyruvate Dehydrogenase E2 on different lysates with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (HA721267) at 1/2,000 dilution.

    Lane 1: Mouse liver tissue lysate
    Lane 2: Rat liver tissue lysate

    Lysates/proteins at 40 µg/Lane.

    Predicted band size: 69 kDa
    Observed band size: 60 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 (HA721267) at 1/2,000 dilution was used in 5% NFDM/TBST 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 Pyruvate Dehydrogenase E2 on different lysates with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (<a href="/products/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Pyruvate Dehydrogenase E2 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 69 kDa<br />Observed band size: 69 kDa<br /><br />Exposure time: 4 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/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/1,000 dilution was used in <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.

    ☑ Knockdown (KD)

    Western blot analysis of Pyruvate Dehydrogenase E2 on different lysates with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (HA721267) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Pyruvate Dehydrogenase E2 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 69 kDa
    Observed band size: 69 kDa

    Exposure time: 4 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 (HA721267) at 1/1,000 dilution was used in 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.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (<a href="/products/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/200 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/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/200 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 human testis tissue with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (HA721267) at 1/200 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 (HA721267) at 1/200 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 cerebellum tissue with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (<a href="/products/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/200 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/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/200 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 cerebellum tissue with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (HA721267) at 1/200 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 (HA721267) at 1/200 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 brain tissue with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (<a href="/products/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/200 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/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/200 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 brain tissue with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (HA721267) at 1/200 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 (HA721267) at 1/200 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 hippocampus tissue with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (<a href="/products/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/200 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/HA721267" style="font-weight: bold;text-decoration: underline;">HA721267</a>) at 1/200 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 hippocampus tissue with Rabbit anti-Pyruvate Dehydrogenase E2 antibody (HA721267) at 1/200 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 (HA721267) at 1/200 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.

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Citation