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ACAT2 Recombinant Rabbit Monoclonal Antibody [JE58-83]

Usd: 385

Specification

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Catalog# HA720032

ACAT2 Recombinant Rabbit Monoclonal Antibody [JE58-83]

Application

  • WB

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

ACAT2 Recombinant Rabbit Monoclonal Antibody [JE58-83]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human ACAT2 aa 198-397/397.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, FC

Target Molecular Weight

Predicted band size: 41 kDa

Positive Control

HeLa cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse liver tissue lysate, Mouse small intestine tissue lysate, Rat liver tissue lysate, A549 cell lysate, human prostate tissue, HepG2.

Conjugation

unconjugated

Clone Number

JE58-83

RRID

AB_3071972

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

  • 1:500-1:2,000

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:500-1:1,000

Target

Function

The product of this gene is an enzyme involved in lipid metabolism, and it encodes cytosolic acetoacetyl-CoA thiolase. This gene shows complementary overlapping with the 3-prime region of the TCP1 gene in both mouse and human. These genes are encoded on opposite strands of DNA, as well as in opposite transcriptional orientation. Alternatively spliced transcript variants encoding different isoforms have been found for this gene.

Background References

1. Weng M. et. al. ACAT2 Promotes Cell Proliferation and Associates with Malignant Progression in Colorectal Cancer. Onco Targets Ther. 2020 Apr

2. Wang YJ. et. al. Cholesterol and fatty acids regulate cysteine ubiquitylation of ACAT2 through competitive oxidation. Nat Cell Biol. 2017 Jul

Subcellular Location

Cytosol.

UNIPROT

SwissProt: Q9BWD1 Human

SwissProt: Q8CAY6 Mouse

SwissProt: Q5XI22 Rat

Synonyms

Acat2 antibody

acetoacetyl CoA thiolase antibody

acetoacetyl Coenzyme A thiolase antibody

Acetyl CoA acetyltransferase antibody

Acetyl CoA transferase like protein antibody

acetyl Coenzyme A acetyltransferase 2 antibody

Acetyl-CoA acetyltransferase antibody

Acetyl-CoA transferase-like protein antibody

Acyl Coenzyme A cholesterol acyltransferase 2 antibody

Cytosolic acetoacetyl-CoA thiolase antibody

Expand

Images

  • Western blot analysis of ACAT2 on different lysates with Rabbit anti-ACAT2 antibody (<a href="/products/HA720032" style="font-weight: bold;text-decoration: underline;">HA720032</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 3: PC-12 cell lysate (20 µg/Lane)<br />Lane 4: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 5: Mouse small intestine tissue lysate (40 µg/Lane)<br />Lane 6: Rat liver tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 41 kDa<br />Observed band size: 41 kDa<br /><br />Exposure time: 14 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/HA720032" style="font-weight: bold;text-decoration: underline;">HA720032</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 ACAT2 on different lysates with Rabbit anti-ACAT2 antibody (HA720032) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 3: PC-12 cell lysate (20 µg/Lane)
    Lane 4: Mouse liver tissue lysate (40 µg/Lane)
    Lane 5: Mouse small intestine tissue lysate (40 µg/Lane)
    Lane 6: Rat liver tissue lysate (40 µg/Lane)

    Predicted band size: 41 kDa
    Observed band size: 41 kDa

    Exposure time: 14 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 (HA720032) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of ACAT2 on different lysates with Rabbit anti-ACAT2 antibody (<a href="/products/HA720032" style="font-weight: bold;text-decoration: underline;">HA720032</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate<br />Lane 2: A549-si ACAT2 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 41 kDa<br />Observed band size: 41 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/HA720032" style="font-weight: bold;text-decoration: underline;">HA720032</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.

    ☑ Knockdown (KD)

    Western blot analysis of ACAT2 on different lysates with Rabbit anti-ACAT2 antibody (HA720032) at 1/2,000 dilution.

    Lane 1: A549-si NT cell lysate
    Lane 2: A549-si ACAT2 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 41 kDa
    Observed band size: 41 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 (HA720032) 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.

  • Immunohistochemical analysis of paraffin-embedded human prostate tissue using anti-ACAT2 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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA720032" style="font-weight: bold;text-decoration: underline;">HA720032</a>, 1/200) 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 human prostate tissue using anti-ACAT2 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 (HA720032, 1/200) 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 ACAT2 was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/HA720032" style="font-weight: bold;text-decoration: underline;">HA720032</a>, 1ug/ml) (red) compared with Rabbit IgG, monoclonal - Isotype Control (green). After incubation of the primary antibody at +4℃ for 1 hour, the cells were stained with a Alexa Fluor&reg;488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃ (dark incubation).Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of ACAT2 was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (HA720032, 1ug/ml) (red) compared with Rabbit IgG, monoclonal - Isotype Control (green). After incubation of the primary antibody at +4℃ for 1 hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃ (dark incubation).Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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