Calpain Activity Fluorometric Assay, Mammalian

Description

The activation of calpain involves many forms of physiological and pathological processes (for example, apoptosis). Calpain activation requires cell membranes and Ca 2+, and activated calpain is released into the cytoplasm. The calpain activity assay kit provides optimized buffers and reagents for easy measurement of calpain activity. The extraction buffer supplied with the kit can specifically extract cytoplasmic proteins without contaminating cell membranes and lysosomal proteases. The extraction buffer can also prevent the automatic activation of calpain during the extraction process. Therefore, after treating the cells with inducers (such as chemicals or drugs), the kit can only detect the activated calpain in the cytosol. The fluorometric method is based on the cleavage detection of the Calpain substrate Ac-LLY-AFC. Ac-LLY-AFC emits blue light (λmax = 400 nm); after calpain cleaves the substrate, free AFC will emit yellow-green fluorescence (λmax = 505 nm), which can be quantified using a fluorometer or a fluorescence microplate reader. Comparing the fluorescence intensity from the treated sample with the normal control can determine the change in calpain activity.
The kit provides a simple and convenient means for analyzing calpain activity in apoptotic and other samples. The assay is based on fluorometric detection of the cleavage of calpain substrate at 505 nm using a fluorometer or a fluorecence plate reader.

Species Reactivity

Mammalian

Relevant Diseases

Neurological Disorders

Detection Method

Fluorescence (Ex/Em 400/505 nm)

Assay Type

ELISA kit

Components

• Extraction Buffer
• 10X Reaction Buffer
• Calpain Substrate Ac-LLY-AFC
• Active Calpain I (Positive Control)
• Calpain Inhibitor Z-LLY-FMK

Shipping

Dry Ice

Storage

-70°C

Research Use Only

For research use only

Target

Calpain

Official Name

Calpain