Cell Signaling Technology

Product Pathways - Autophagy Signaling

Chloroquine #14774

autophagy   GABARAP   LC3   malarial  

Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype

Species cross-reactivity is determined by western blot.

Applications Key:

Description

Molecular Weight:

515.9 g/mol

Molecular Characterization

C18H26ClN3•2H3PO4

Purity

>98%

Bioactivity

Solubility: Soluble in water at 50 mg/ml. Poorly soluble in DMSO and ethanol.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated (-) or treated with Chloroquine (50 μM, overnight; +), using LC3A/B (D3U4C) XP® Rabbit mAb #12741.

Structure

Structure

Chemical structure of chloroquine.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa (upper) and C2C12 (lower) cells, treated with Chloroquine (50 μM, overnight; left), nutrient-starved with EBSS (3 hr; middle), or untreated (right), using LC3A/B (D3U4C) XP® Rabbit mAb #12741 (green) and β-Actin (13E5) Rabbit mAb (Alexa Fluor® 555 Conjugate) #8046 (red). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).

Directions for Use

Chloroquine is supplied as a lyophilized powder. For a 50 mM stock, reconstitute the 150 mg in 5.82 ml sterile dH2O. First add 1 ml dH2O to the tube containing the chemical, vortex, and dispense into a new, larger tube. Repeat this action two or three more times to transfer any residual material. Add additional dH2O to the new tube to bring the volume up to 5.82 ml. Filter sterilize into sterile tube. Utilize a syringe and 0.2 μm syringe filter to minimize sample loss.

Working concentrations and length of treatment can vary depending on the desired effect, but it is typically used at 25-100 μM for 12-48 hr.

Background

Chloroquine (CQ) is a lysosomotropic agent with an extensive range of biological effects (1). Historically known for its anti-malarial activity, chloroquine is a widely used biological research tool for studying autophagy inhibition. Research studies demonstrate that chloroquine accumulates in acidic lysosomes and increases the lysosomal pH. This inhibits lysosomal hydrolases and prevents autophagosomal fusion and degradation, which can result in apoptotic or necrotic cell death (1-4). Inhibition of chloroquine-induced apoptosis with the V-ATPase inhibitor bafilomycin A1 has been observed in several cell types (4). Chloroquine also enhances the anti-neoplastic effects of the histone deacetylase inhibitor vorinostat (SAHA) (5).

Chloroquine treatment of cells leads to accumulation of light chain 3-II (LC3-II) (1-3). This autophagy marker resides within autophagosomal membranes during the autophagic process and is degraded upon fusion with lysosomes. Chloroquine inhibition of these fusion events effectively blocks LC3-II degradation.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products


For Research Use Only. Not For Use In Diagnostic Procedures.

DRAQ5 is a registered trademark of Biostatus Limited.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

用户评论 --- 共 0

该产品暂无评论!

我要参与评论 :

如要参与评论请先登录网站

还没有网站账户?去注册一下吧

Products

 

Applications