HeLa Mitochondrial DNA Depletion Cell Lysate Set: Ethidium Bromide-Treated and Vehicle-Treated (ab154479)
Overview
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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/mLHuman 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
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Tested applications
Suitable for: WB, SDS-PAGEmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Storage buffer
Constituents: 0.03% Sodium fluoride, 0.01% Bromophenol blue, 1.8% SDS, 0.77% DTT, 1.1% Tris, 10% Glycerol, 0.7% Sodium chloride -
Components 2 units Control for Ethidium Bromide-Treated HeLa Lysate 1 x 200µg Ethidium Bromide-Treated HeLa Lysate 1 x 200µg -
Research areas
Images
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Western blot - Rho0 Whole Cell Lysate (ab154479)
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, 15µgBlock 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 -
SDS-PAGE - Rho0 Whole Cell Lysate (ab154479)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