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ACO1 Recombinant Rabbit Monoclonal Antibody [JE63-98]

Usd: 385

Specification

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Catalog# HA722071

ACO1 Recombinant Rabbit Monoclonal Antibody [JE63-98]

Application

  • WB

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

ACO1 Recombinant Rabbit Monoclonal Antibody [JE63-98]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human ACO1 aa 1-50 / 889.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, FC

Molecular Weight

Predicted band size: 98 kDa

Positive Control

HeLa cell lysate, 293T cell lysate, HCT 116 cell lysate, mouse liver tissue lysate, rat liver tissue lysate, HeLa.

Conjugation

unconjugated

Clone Number

JE63-98

RRID

AB_3096724

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

  • 1:2,000-1:5,000

  • FC

  • 1:1,000

Target

Function

Iron metabolism is essential for sustaining mammalian homeostasis. Iron uptake and distribution is a highly regulated process in mammalian cells that is monitored by two iron sensing proteins; iron regulatory protein-1 and -2 (IRP-1 and -2), also known as iron responsive element-binding protein-1 and -2 (IRE-BP-1 and -2) or aconitase 1 and 2. IRP-1 and IRP-2 are important soluble regulatory factors that mediate iron uptake and storage in mammalian cells. They are capable of either repressing translation or enhancing mRNA stability by associating with stem-loop motifs known as iron-responsive elements (IREs). IRPs respond to stress mediators, iron concentration and signaling factors, including nitrogen monoxide, cytokines and hydrogen peroxide.

Background References

1. Metodiev MD et al. Mutations in the tricarboxylic acid cycle enzyme, aconitase 2, cause either isolated or syndromic optic neuropathy with encephalopathy and cerebellar atrophy. J Med Genet 51:834-8 (2014).

2. Jiang X et al. Hyperinsulinemia induces hepatic iron overload by increasing liver TFR1 via the PI3K/IRP2 pathway. J Mol Endocrinol 53:381-92 (2014).

Subcellular Location

Cytoplasm, cytosol.

UNIPROT

SwissProt: P21399 Human

SwissProt: P28271 Mouse

SwissProt: Q63270 Rat

Synonyms

ACO 1 antibody

ACO1 antibody

ACOC_HUMAN antibody

Aconitase 1 soluble antibody

Aconitase antibody

Aconitase1 antibody

Aconitate hydratase antibody

ACONS antibody

Citrate hydro lyase antibody

Citrate hydro-lyase antibody

Expand

Images

  • Western blot analysis of ACO1 on different lysates with Rabbit anti-ACO1 antibody (<a href="/products/HA722071" style="font-weight: bold;text-decoration: underline;">HA722071</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: 293T cell lysate (20 µg/Lane)<br />Lane 3: HCT 116 cell lysate (20 µg/Lane)<br />Lane 4: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 5: Rat liver tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 98 kDa<br />Observed band size: 98 kDa<br /><br />Exposure time: 5 minutes 10 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/HA722071" style="font-weight: bold;text-decoration: underline;">HA722071</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 ACO1 on different lysates with Rabbit anti-ACO1 antibody (HA722071) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: 293T cell lysate (20 µg/Lane)
    Lane 3: HCT 116 cell lysate (20 µg/Lane)
    Lane 4: Mouse liver tissue lysate (40 µg/Lane)
    Lane 5: Rat liver tissue lysate (40 µg/Lane)

    Predicted band size: 98 kDa
    Observed band size: 98 kDa

    Exposure time: 5 minutes 10 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 (HA722071) 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 ACO1 on different lysates with Rabbit anti-ACO1 antibody (<a href="/products/HA722071" style="font-weight: bold;text-decoration: underline;">HA722071</a>) at 1/5,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-ACO1 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 98 kDa<br />Observed band size: 98 kDa<br /><br />Exposure time: 30 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/HA722071" style="font-weight: bold;text-decoration: underline;">HA722071</a>) at 1/5,000 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 ACO1 on different lysates with Rabbit anti-ACO1 antibody (HA722071) at 1/5,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-ACO1 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 98 kDa
    Observed band size: 98 kDa

    Exposure time: 30 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 (HA722071) at 1/5,000 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.

  • Flow cytometric analysis of HeLa cells labeling ACO1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722071" style="font-weight: bold;text-decoration: underline;">HA722071</a>, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling ACO1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722071, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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