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ACAT-1 Recombinant Rabbit Monoclonal Antibody [JE64-40]

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Specification

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Catalog# HA721093

ACAT-1 Recombinant Rabbit Monoclonal Antibody [JE64-40]

Application

  • WB

  • IHC-P

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

ACAT-1 Recombinant Rabbit Monoclonal Antibody [JE64-40]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human ACAT1 aa 378-427/427.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell

Molecular Weight

Predicted band size: 45 kDa

Positive Control

Raji cell lysate, HepG2 cell lysate, rat liver tissue lysate, rat heart tissue lysate, HepG2, human liver tissue, human liver carcinoma tissue, rat liver tissue, mouse liver tissue, mouse heart tissue.

Conjugation

unconjugated

Clone Number

JE64-40

RRID

AB_3072217

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

  • 1:500-1:1,000

  • IHC-P

  • 1:100-1:400

  • IF-Cell

  • 1:250

Target

Function

Acetyl-CoA acetyltransferase, mitochondrial, also known as acetoacetyl-CoA thiolase, is an enzyme that in humans is encoded by the ACAT1 (Acetyl-Coenzyme A acetyltransferase 1) gene. Acetyl-Coenzyme A acetyltransferase 1 is an acetyl-CoA C-acetyltransferase enzyme. This gene encodes a mitochondrially localized enzyme that catalyzes the reversible formation of acetoacetyl-CoA from two molecules of acetyl-CoA. The ACAT1 enzyme has a few unique properties. First, it is activated by potassium ions binding near the CoA binding site and the catalytic site. This binding causes a structural change in the active site loop. Additionally, this enzyme is able to use 2-methyl-branched acetoacetyl-CoA as a substrate, making it a unique thiolase. ACAT1 is regulated at both transcriptional and translational levels. ACAT1 enzyme activity is enhanced ACAT1’s expression is promoted transcriptionally by leptin, angiotensin II, and insulin in human monocytes/macrophages. Insulin-mediated regulation also involves ERK, p38MAPK, and JNK signaling pathways.

Background References

1. Gu L. et. al. Stabilization of FASN by ACAT1-mediated GNPAT acetylation promotes lipid metabolism and hepatocarcinogenesis. Oncogene. 2020 Mar

2. Qian H. et. al. Structural basis for catalysis and substrate specificity of human ACAT1. Nature. 2020 May

Subcellular Location

Mitochondrion.

UNIPROT

SwissProt: P24752 Human

SwissProt: Q8QZT1 Mouse

SwissProt: P17764 Rat

Synonyms

ACAT 1 antibody

ACAT antibody

acat1 antibody

Acetoacetyl CoA thiolase antibody

acetoacetyl Coenzyme A thiolase antibody

Acetoacetyl-CoA thiolase antibody

Acetyl CoA acetyltransferase, mitochondrial antibody

Acetyl Coenzyme A acetyltransferase 1 antibody

Acetyl-CoA acetyltransferase antibody

acetyl-coa acetyltransferase precursor, mitochondrial antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of ACAT-1 on different lysates with Rabbit anti-ACAT-1 antibody (<a href="/products/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate<br />Lane 2: A549-si ACAT-1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 45 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 1 minute; 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/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</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 ACAT-1 on different lysates with Rabbit anti-ACAT-1 antibody (HA721093) at 1/2,000 dilution.

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

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 45 kDa
    Observed band size: 42 kDa

    Exposure time: 1 minute; 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 (HA721093) 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.

  • Western blot analysis of ACAT-1 on different lysates with Rabbit anti-ACAT-1 antibody (<a href="/products/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/500 dilution.<br /><br />Lane 1: Raji cell lysate<br />Lane 2: HepG2 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 45 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 4 minutes;<br /><br />12% 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/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:200,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of ACAT-1 on different lysates with Rabbit anti-ACAT-1 antibody (HA721093) at 1/500 dilution.

    Lane 1: Raji cell lysate
    Lane 2: HepG2 cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 45 kDa
    Observed band size: 42 kDa

    Exposure time: 4 minutes;

    12% 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 (HA721093) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of ACAT-1 on different lysates with Rabbit anti-ACAT-1 antibody (<a href="/products/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/500 dilution.<br /><br />Lane 1: Rat liver tissue lysate<br />Lane 2: Rat heart tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 45 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 1 minutes;<br />8% 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/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of ACAT-1 on different lysates with Rabbit anti-ACAT-1 antibody (HA721093) at 1/500 dilution.

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

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 45 kDa
    Observed band size: 42 kDa

    Exposure time: 1 minutes;
    8% 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 (HA721093) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HepG2 cells labeling ACAT-1 with Rabbit anti-ACAT-1 antibody (<a href="/products/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/250 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ACAT-1 antibody (<a href="/products/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of HepG2 cells labeling ACAT-1 with Rabbit anti-ACAT-1 antibody (HA721093) at 1/250 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ACAT-1 antibody (HA721093) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Counterstained with Mitotracker. Nuclear DNA was labelled in blue with DAPI.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-ACAT-1 antibody (<a href="/products/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</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/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</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 liver tissue with Rabbit anti-ACAT-1 antibody (HA721093) 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 (HA721093) 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 liver carcinoma tissue with Rabbit anti-ACAT-1 antibody (<a href="/products/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</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/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</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 liver carcinoma tissue with Rabbit anti-ACAT-1 antibody (HA721093) 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 (HA721093) 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 liver tissue with Rabbit anti-ACAT-1 antibody (<a href="/products/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/100 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/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/100 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 liver tissue with Rabbit anti-ACAT-1 antibody (HA721093) at 1/100 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 (HA721093) at 1/100 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 liver tissue with Rabbit anti-ACAT-1 antibody (<a href="/products/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/100 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/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/100 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 liver tissue with Rabbit anti-ACAT-1 antibody (HA721093) at 1/100 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 (HA721093) at 1/100 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-ACAT-1 antibody (<a href="/products/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/400 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/HA721093" style="font-weight: bold;text-decoration: underline;">HA721093</a>) at 1/400 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-ACAT-1 antibody (HA721093) at 1/400 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 (HA721093) at 1/400 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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