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Autophagy Control: HeLa Chloroquine-Treated and Control Lysate (ab171461) is designed for use as a Western blot control when studying protein extracts for autophagic activity. Cells were treated overnight with chloroquine in the presence of serum. Untreated cells were grown under standard conditions. Treatment induces increase in LC3B II in comparison to the untreated control.
Chloroquine is a lysosomotropic compound that inhibits lysosomal acid hydrolases by raising lysosomal pH. The change in pH prevents lysosomal degradation of autophagosomes and therefore accumulation of autophagosomal membranes.
Concentration: HeLa Chloroquine-treated lysate, 200 µg at 2.0 mg/mL
HeLa DMSO-treated lysate, 200 µg at 2.0 mg/mL
|HeLa Chloroquine-treated lysate||1 x 200µg|
|HeLa control lysate||1 x 200µg|
Our Abpromise guarantee covers the use of ab171461 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration.
Load 20 µg – 40 µg /lane. Heat sample at 70°C before loading.
|SDS-PAGE||Use at an assay dependent concentration.|
HeLa cell cultures were treated with 50 µM chloroquine overnight at 37°C. 40 µg cell lysate was run on a 4-20% gradient gel then stained with Optiblot Blue (ab119211) for 2 hours at room temperature. Gel was destained using nanopure water for 24 hours.
HeLa cell cultures were treated with 50 µM chloroquine overnight at 37°C. 40 µg cell lysate was run on a 4-20% gradient gel then transferred to a PVDF membrane. All blocking and antibody incubation steps were done in 5% milk, 20 mM Tris-HCl, 0.1% TWEEN-20. Primary antibody: ab51520 used at 1/3000. Secondary antibody: Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/10000. Observed bands: LC3B-I: 16 kDa and LC3B-II: 14 kDa.
ab171461 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"