Invitrogen™ CellEvent™ Senescence Green Detection Kit

Description

• This fluorescein-based probe contains two galactoside moieties, making it a target for β-galactosidase
• Activation of β-galactosidase is commonly used as a biomarker for senescent cells
• This hydrolase enzyme resides in lysosomes and converts β-galactosides into monosaccharides under acidic pH conditions
• The enzymatically cleaved product is retained within the cell due to covalent binding of intracellular proteins and emits a fluorogenic signal that has absorption/emission maxima of 490/514 nm
• Kit features include: • Reliable and quick fluorescent detection of senescent cells • Greater sensitivity than traditional colorimetric x-gal • Fluorescent signal is well retained and detected using standard Alexa Fluor™ 488/FITC filter sets • Multiplex-enabled—fluorescent senescence probe can be multiplexed Due to a limited replicative lifespan, normal cells enter cell cycle arrest, also known as cellular senescence
• While in this senescent phase the cells remain metabolically active without undergoing cell death or division
• They adopt a specific phenotypic that includes the appearance of multiple nuclei, increased vacuolization, expression of pH-dependent β-galactosidase, and morphological enlargement and extension
• Senescence, through a variety of mechanisms, may play a role in tumor suppression, tumor progression, aging, and tissue repair
• Activation of β-galactosidase is commonly used as a biomarker for senescent cells
• This hydrolase enzyme resides in lysosomes and converts β-galactosides into monosaccharides under acidic pH conditions
• The activity is optimal at lysosomal pH 4, but conventional assays measure at pH 6
• It has been shown that normalized β-galactosidase activity is twice as high in senescent cells as in pre-senescent cells regardless of the pH value used for testing
• A colorimetric substrate for β-galactosidase, 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (x-gal), has long been used to detect metabolic activity in cells in vitro
• However, its use is limited due to inconsistent signal, length of assay, and inability to multiplex
• Therefore, we have developed a sensitive, fluorescent substrate for β-galactosidase that can be used for the detection of senescent cells
• The CellEvent™ Senescence Green probe is a fluorescein-based reagent that contains two galactoside moieties, making it a specific target of β-galactosidase
• The enzyme- cleaved product is retained within the cell due to covalent binding of intracellular proteins and emits a fluorogenic signal that has absorption/emission maxima of 490/514 nm
• The CellEvent™ Senescent Green Kit provides CellEvent™ Senescent Green probe and an optimized buffer for the detection of senescent cells in fixed samples
• This probe can be multiplexed with other fluorescent reagents compatible with paraformaldehyde fixation.