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Invitrogen™ CellEvent™ Senescence Green Flow Cytometry Assay Kit
Description
The kit contains an optimized buffer and a fluorescein-based reagent that contains two galactoside moieties, making it a specific 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 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.
CellEvent Senescence Green Flow Cytometry Assay Kit features include:
• Fluorescent detection of β-galactosidase hydrolysis by flow cytometry
• Fixed cell assay
• Multi-color compatibility
When normal cells reach the end of their limited replicative lifespan, commonly referred to as the Hayflick limit, they enter into a senescent phase where they remain metabolically active without undergoing cell death processes. These senescent cells adopt a specific phenotypic state that includes the appearance of multi-nucleated cells, increased vacuolization, expression of pH-dependent β-galactosidase, and morphological changes where cells become enlarged and extended. Through a variety of mechanisms, they may play a role in tumor suppression, tumor progression, aging, and tissue repair.
β-galactosidase resides in lysosomes and converts β-galactosides into monosaccharides under acidic pH conditions. The activity is optimal under 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 presenescent cells regardless of the pH value used for testing.
Traditionally, the 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. Upon hydrolysis by β-galactosidase, x-gal converts to a blue precipitate, which is not suitable for flow cytometric assays. Therefore, we have developed a sensitive, fluorescent substrate for β-galactosidase that can be used in flow cytometry assays for the detection of senescent cells.
Specifications
Specifications
| Color | Green |
| Content And Storage | Store at 2°C to 8°C and protect from light. |
| Detection Method | Fluorescence |
| For Use With (Application) | Flow Cytometry |
| For Use With (Equipment) | Flow Cytometer |
| Product Type | Flow Cytometry Assay Kit |
| Emission | 488/514 |
| Product Line | CellEvent |
| Quantity | 50 Assays |
| Shipping Condition | Room Temperature |
Frequently Asked Questions (FAQs)
We do not recommend skipping the fixation step when using CellEvent Senescence Green Flow Cytometry Assay Kit, as it is a necessary step. The CellEvent Senescence Green probe only works on fixed cells.
Yes. After incubating the cells with CellEvent Senescence Green Probe, permeabilize the cells, wash, and proceed with detection.
We do not recommend using your own buffer with the CellEvent Senescence Green Probe. The assay is pH sensitive and we recommend that you use the buffer provided in the kit be used.
No. It only works on fixed cells; b-galactosidase is still active immediately following fixation with paraformaldehyde.
For Research Use Only. Not for use in diagnostic procedures.
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