HeLa Mitochondrial DNA Depletion Cell Lysate Set: Ethidium Bromide-Treated and Vehicle-Treated (ab154479)

Overview

  • Product name
    HeLa Mitochondrial DNA Depletion Cell Lysate Set: Ethidium Bromide-Treated and Vehicle-Treated
  • Product overview

    HeLa cells were cultured in the presence or absence of 50 ng/mL Ethidium Bromide for 14 days. Whole cell lysates were prepared in an SDS lysis buffer containing protease and phosphatase inhibitors. Ethidium bromide cells are judged to be depleted of mitochondrial DNA (termed Rho0) on the basis of western blotting.

     

    Concentration: HeLa Rho0 lysate, 200 µg at 1.0 mg/mL

                                HeLa control lysate, 200 µg at 1.0 mg/mL

     

    Human mitochondrial DNA (mtDNA) encodes 13 subunits of the oxidative phosphorylation (OXPHOS) enzyme complexes I, III, IV and V. Low concentrations of ethidium bromide inhibit mtDNA synthesis without having overtly effecting cell viability. Loss of mtDNA not only depletes mtDNA encoded proteins but also can lead to decreased levels of nuclear DNA encoded mitochondrial proteins due to protein complex instabilities

  • Tested applications
    Suitable for: WB, SDS-PAGEmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab154479 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration.
SDS-PAGE Use at an assay dependent concentration. Customer may choose to boil the sample before loading, at their discretion.

Images

  • Western blot of HeLa Whole Cell Lysate: Rho treated and control (ab154479) using MitoProfile® Total OXPHOS Human WB Antibody Cocktail (ab110411). The results show a partial or complete reduction of subunits across the five OXPHOS complexes 1-V.

    Lane 1 HeLa control lysate, 15µg
    Lane 2 HeLa Rho0 lysate, 15µg

    Block with 4% milk/PBS
    Primary antibody: MitoProfile® Total OXPHOS Human WB Antibody Cocktail (ab110411), 1/208.
    Secondary antibody: anti-mouse-HRP, 1/5000
    ECL detection

  • Coomassie blue stained SDS-PAGE gel of HeLa Control and Rho0 lysates (ab154479) to demonstrate equivalent protein normalization.

    Lane1. MW Markers
    Lane2. HeLa control lysate (ab154479); 20µg load
    Lane3. HeLa Rho0 lysate (ab154479); 20µg load

References

ab154479 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

The kit contains 200ug of Hela control lysate and 200ug of HeLa Rho0 lysate.

The concentration of both lysates is at 1mg/mL.

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

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