Cell Proliferation Tracer Kit, (Fluorometric, Blue 520) #53452
Live human peripheral blood mononuclear cells were labeled with Cell Proliferation Tracer Kit (Fluorometric, Blue 520). Cells cultured in medium containing Human Interleukin-2 (hIL-2) #8907 (50 ng/mL) were then left untreated (blue solid line) or stimulated with anti-CD3 (10 μg/mL) and anti-CD28 (5 μg/ml) (green solid line). Cells were analyzed by flow cytometry after 72 hr. Unstained cells were also acquired for comparison (blue dashed line).
- Product Includes
- Related Products
| Product Includes | Quantity (with Count) |
|---|---|
| Cell Proliferation Tracer Dye, Blue 520 #73595 | 1 x 1 ea |
| Anhydrous DMSO #15360 | 1 x 150 µl |
Product Information
Protocol
Product Description
The Cell Proliferation Tracer Kit (Fluorometric, Blue 520) contains Cell Proliferation Tracer Dye, Blue 520 that diffuses passively into live cells and is used for long-term cell labeling. This dye is initially a non-fluorescent ester, but is converted to a fluorescent dye by intracellular esterases. The dye then covalently reacts with amine groups on proteins, forming fluorescent conjugates that are retained in the cell. Immediately after staining, a single, bright fluorescent population will be detected by flow cytometry. Each cell division that occurs after labeling results in the appearance of a dimmer fluorescent peak on a flow cytometry histogram. The Cell Proliferation Tracer Kit (Fluorometric, Blue 520) can be used to track cell divisions in vivo or in vitro. Staining can withstand fixation and permeabilization for subsequent immunostaining.
Background
Due to their inherent metabolic stability once inside a cell, fluorescent proliferation dyes partition in an equal manner between daughter cells during the M phase of the cell cycle. This allows the principle of dye dilution to be leveraged as a means to trace multiple rounds of cell proliferation using flow cytometry. Added benefits of proliferation dyes are that they are non-radioactive and do not require cells to be actively synthesizing DNA for efficient uptake (1-3).
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