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Fatty Acid and Lipid Metabolism Antibody Sampler Kit

Usd: 640

Specification

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Catalog# HAK21048

Fatty Acid and Lipid Metabolism Antibody Sampler Kit

Application

  • WB

Reactivity

  • Human

  • Mouse

Overview

Kit Components

Product Includes Specification Application Reactivity Mw
Acetyl CoA synthetase[ET1702-21] 20µl WB,IF-Cell,IF-Tissue Human,Mouse,Rat Predicted band size: 79 kDa
Acetyl CoA Carboxylase 1 (ACC1)[ET1609-77] 20µl WB,IHC-P Human,Mouse,Rat Predicted band size: 266 kDa
ATP citrate lyase[ET1609-37] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC,IP Human,Mouse,Rat Predicted band size: 121 kDa
Fatty Acid Synthase[ET1701-91] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC,IP Human,Mouse,Rat Predicted band size: 273 kDa
Lipin 1[ET7108-51] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC,IP Human,Mouse 98 kDa
ACSL1[HA601112] 20µl WB,IHC-P Human,Mouse,Rat Predicted band size: 78 kDa
Phospho-Acetyl Coenzyme A Carboxylase (S79)[HA721714] 20µl WB Human,Mouse,Rat Predicted band size: 265 kDa
Goat Anti-Rabbit IgG (H+L)[HA1001] 100µl WB,ELISA,IHC-P Rabbit
Goat Anti-Mouse IgG (H+L)[HA1006] 100µl WB,ELISA,IHC-P Mouse

Product Description

The Fatty Acid and Lipid Metabolism Antibody Sampler Kit provides an economical means to evaluate key proteins involved in fatty acid and lipid metabolism. This kit includes enough primary antibody to perform two western miniblot experiments with each primary antibody.

Product Features

Storage Buffer

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

Storage Instructions

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

Background

The processes of fatty acid and lipid metabolism are vital for cellular nutrient and energy maintenance. Cytoplasmic acetyl-CoA synthetase (AceCS1) catalyzes the conversion of acetate and CoA to acetyl-CoA. Acetyl-CoA carboxylase (ACC) catalyzes the pivotal step of the fatty acid synthesis pathway. <BR>Phosphorylation by AMPK at Ser79 or by PKA at Ser1200 inhibits the enzymatic activity of ACC. ATP-citrate lyase (ACL) is a homotetramer that catalyzes the formation of acetyl-CoA and oxaloacetate (OAA) in the cytosol, which is the key step for the biosynthesis of fatty acids, cholesterol, and acetylcholine, as well as for glucogenesis. Phosphorylation of ACL at Ser455 abolishes the homotropic allosteric regulation by citrate and enhances the catalytic activity of the enzyme. Fatty acid synthase (FASN) catalyzes the synthesis of long-chain fatty acids from acetyl-CoA and malonyl-CoA. Lipin 1 plays a role in lipid metabolism in various tissues and cell types including liver, muscle, adipose tissues, and neuronal cell lines.

Data Links

Background References

1. Reue K, Zhang P. The lipin protein family: dual roles in lipid biosynthesis and gene expression. FEBS Lett. 2008 Jan 9;582(1):90-6.

2. Li C, Zhang L, Qiu Z, Deng W, Wang W. Key Molecules of Fatty Acid Metabolism in Gastric Cancer. Biomolecules. 2022 May 15;12(5):706.

Images

  • Western blot analysis of Acetyl CoA synthetase on different lysates with Rabbit anti-Acetyl CoA synthetase antibody (<a href="/products/ET1702-21" style="font-weight: bold;text-decoration: underline;">ET1702-21</a>) at 1/1,000 dilution.<br /><br />Lane 1: HepG2 cell lysate (20 µg/Lane)<br />Lane 2: L6 cell lysate (20 µg/Lane)<br />Lane 3: COS-1 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 79 kDa<br />Observed band size: 79 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/ET1702-21" style="font-weight: bold;text-decoration: underline;">ET1702-21</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 Acetyl CoA synthetase on different lysates with Rabbit anti-Acetyl CoA synthetase antibody (ET1702-21) at 1/1,000 dilution.

    Lane 1: HepG2 cell lysate (20 µg/Lane)
    Lane 2: L6 cell lysate (20 µg/Lane)
    Lane 3: COS-1 cell lysate (20 µg/Lane)

    Predicted band size: 79 kDa
    Observed band size: 79 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 (ET1702-21) 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 Acetyl CoA Carboxylase 1 (ACC1) on different lysates with Rabbit anti-Acetyl CoA Carboxylase 1 (ACC1) antibody (<a href="/products/ET1609-77" style="font-weight: bold;text-decoration: underline;">ET1609-77</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: HepG2 cell lysate (20 µg/Lane)<br />Lane 3: HEK-293 cell lysate (20 µg/Lane)<br />Lane 4: A549 cell lysate (20 µg/Lane)<br />Lane 5: C2C12 cell lysate (20 µg/Lane)<br />Lane 6: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 7: C6 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 266 kDa<br />Observed band size: 250/266 kDa<br /><br />Exposure time: 43 seconds;<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/ET1609-77" style="font-weight: bold;text-decoration: underline;">ET1609-77</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 Acetyl CoA Carboxylase 1 (ACC1) on different lysates with Rabbit anti-Acetyl CoA Carboxylase 1 (ACC1) antibody (ET1609-77) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: HepG2 cell lysate (20 µg/Lane)
    Lane 3: HEK-293 cell lysate (20 µg/Lane)
    Lane 4: A549 cell lysate (20 µg/Lane)
    Lane 5: C2C12 cell lysate (20 µg/Lane)
    Lane 6: RAW264.7 cell lysate (20 µg/Lane)
    Lane 7: C6 cell lysate (20 µg/Lane)

    Predicted band size: 266 kDa
    Observed band size: 250/266 kDa

    Exposure time: 43 seconds;

    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 (ET1609-77) 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 ATP citrate lyase with anti-ATP citrate lyase antibody [ST51-07] (<a href="/products/ET1609-37" style="font-weight: bold;text-decoration: underline;">ET1609-37</a>) at 1/1,000 dilution.<br />Lane 1: Wild-type A549 whole cell lysate (20 µg).<br />Lane 2: ATP citrate lyase knockout A549 whole cell lysate (20 µg).<br /><br /><a href="/products/ET1609-37" style="font-weight: bold;text-decoration: underline;">ET1609-37</a> was shown to specifically react with ATP citrate lyase in wild-type A549 cells. No band was observed when ATP citrate lyase knockout sample was tested. Wild-type and ATP citrate lyase knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1609-37" style="font-weight: bold;text-decoration: underline;">ET1609-37</a>, 1/1,000) and Loading control antibody (Rabbit anti-HSP90, <a href="/products/ET1605-56" style="font-weight: bold;text-decoration: underline;">ET1605-56</a>, 1/10,000)was used in 5% BSA 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 ATP citrate lyase with anti-ATP citrate lyase antibody [ST51-07] (ET1609-37) at 1/1,000 dilution.
    Lane 1: Wild-type A549 whole cell lysate (20 µg).
    Lane 2: ATP citrate lyase knockout A549 whole cell lysate (20 µg).

    ET1609-37 was shown to specifically react with ATP citrate lyase in wild-type A549 cells. No band was observed when ATP citrate lyase knockout sample was tested. Wild-type and ATP citrate lyase knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1609-37, 1/1,000) and Loading control antibody (Rabbit anti-HSP90, ET1605-56, 1/10,000)was used in 5% BSA 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 Fatty Acid Synthase on different lysates with Rabbit anti-Fatty Acid Synthase antibody (<a href="/products/ET1701-91" style="font-weight: bold;text-decoration: underline;">ET1701-91</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HEK-293 cell lysate<br />Lane 3: A549 cell lysate<br />Lane 4: C2C12 cell lysate<br />Lane 5: L-929 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 273 kDa<br />Observed band size: 273 kDa<br /><br />Exposure time: 1 minute 2 seconds;<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/ET1701-91" style="font-weight: bold;text-decoration: underline;">ET1701-91</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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 Fatty Acid Synthase on different lysates with Rabbit anti-Fatty Acid Synthase antibody (ET1701-91) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HEK-293 cell lysate
    Lane 3: A549 cell lysate
    Lane 4: C2C12 cell lysate
    Lane 5: L-929 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 273 kDa
    Observed band size: 273 kDa

    Exposure time: 1 minute 2 seconds;

    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 (ET1701-91) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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 ACSL1 on different lysates with Mouse anti-ACSL1 antibody (<a href="/products/HA601112" style="font-weight: bold;text-decoration: underline;">HA601112</a>) at 1/1,000 dilution.<br /><br />Lane 1: Raji cell lysate (10 µg/Lane)<br />Lane 2: HepG2 cell lysate (10 µg/Lane)<br />Lane 3: LO2 cell lysate (10 µg/Lane)<br />Lane 4: A431 cell lysate (10 µg/Lane)<br /><br />Predicted band size: 78 kDa<br />Observed band size: 78 kDa<br /><br />Exposure time: 2 minutes;<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/HA601112" style="font-weight: bold;text-decoration: underline;">HA601112</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:150,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of ACSL1 on different lysates with Mouse anti-ACSL1 antibody (HA601112) at 1/1,000 dilution.

    Lane 1: Raji cell lysate (10 µg/Lane)
    Lane 2: HepG2 cell lysate (10 µg/Lane)
    Lane 3: LO2 cell lysate (10 µg/Lane)
    Lane 4: A431 cell lysate (10 µg/Lane)

    Predicted band size: 78 kDa
    Observed band size: 78 kDa

    Exposure time: 2 minutes;

    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 (HA601112) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Phospho-Acetyl Coenzyme A Carboxylase (S79) on different lysates with Rabbit anti-Phospho-Acetyl Coenzyme A Carboxylase (S79) antibody (<a href="/products/HA721714" style="font-weight: bold;text-decoration: underline;">HA721714</a>) at 1/2,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate<br />Lane 2: NIH/3T3 treated with 0.5μM Oligomycin for 30 minutes cell lysate<br />Lane 3: NIH/3T3 treated with 0.5μM Oligomycin for 30 minutes then treated with λpp for 1 hour cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 265 kDa<br />Observed band size: 265 kDa<br /><br />Exposure time: 1 minutes 14 seconds;<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/HA721714" style="font-weight: bold;text-decoration: underline;">HA721714</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 Phospho-Acetyl Coenzyme A Carboxylase (S79) on different lysates with Rabbit anti-Phospho-Acetyl Coenzyme A Carboxylase (S79) antibody (HA721714) at 1/2,000 dilution.

    Lane 1: NIH/3T3 cell lysate
    Lane 2: NIH/3T3 treated with 0.5μM Oligomycin for 30 minutes cell lysate
    Lane 3: NIH/3T3 treated with 0.5μM Oligomycin for 30 minutes then treated with λpp for 1 hour cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 265 kDa
    Observed band size: 265 kDa

    Exposure time: 1 minutes 14 seconds;

    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 (HA721714) 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.

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

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