Live/Dead Cell Staining Kit [for Cell Staining]

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Size	Unit Price	Philadelphia, PA	Portland, OR	Japan*	Quantity	Shipping Information
1KIT	$321.00	Contact Us	Contact Us	13		Shipping Simulation

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Product Information

• Components : Calcein-AM Solution (Ex/Em = 490 nm/515 nm), PI Solution (Ex/Em = 530 nm/620 nm)
• Size : Calcein-AM Solution (1 mM) in DMSO50 µL×4 vials, PI Solution (1.5 mM) in Water600 µL×1 vial

Description

• This product is a combination of Calcein-AM, a fluorescent dye for staining live cells, and PI (Propidium Iodide), a fluorescent dye for staining dead cells, allowing simultaneous staining of live and dead cells. It can be used to observe cells under a fluorescence microscope.

Direction for Use

[Staining Solution Preparation]

• 1. Bring Calcein-AM solution and PI solution to room temperature.
  2. Add 1 μL Calcein-AM solution and 3 μL PI solution to 1mL PBS. This is the staining solution. At this time, the concentrations of Calcein-AM and PI are 1 μM and 4.5 μM, respectively.
• Note: Staining conditions will vary depending on observation conditions such as cell type and concentration. Optimal conditions such as reagent concentration should be considered.
  Calcein-AM can be hydrolyzed by moisture, so replace the cap tightly after use and be careful not to absorb moisture.
  Prepared staining solutions should be used on the same day.
  PI is a suspected carcinogen and should be handled with caution.

[Determination of Optimal Staining Concentration]

• Kill cells by treating with 0.1% saponin or 0.1-0.5% digitonin for 10 minutes or 70% methanol for 30 minutes.
  Stain dead cells with 0.1-10 μM PI solution to find a concentration that stains only the nuclei red without staining the cytoplasm.
  Stain dead cells with 0.1-10 μM Calcein-AM solution to find a concentration that does not stain the cytoplasm. Then confirm that live cells stain at this concentration. If staining is inadequate, increase the concentration.

[Cell Staining]

• 1. For adherent cells, digest cells with cell scraper or trypsin-EDTA and collect by centrifugation (1000 rpm, 3 minutes). For suspended cells, collect cells by direct centrifugation (1000 rpm, 3 minutes).
  2. Centrifuge the cell suspension, remove the supernatant and add PBS. Thoroughly disperse the cells by pipetting or other means. Repeat washing with PBS and centrifugation 2-3 times. Cells should be washed prior to testing, as esterases present in serum containing medium may hydrolyze Calcein-AM resulting in increased background.
  3. After centrifugation, remove the supernatant and add the staining solution (adjust cell count to approximately 0.5 x10⁵ - 10 x10⁵ cells/mL).
  4. Incubate at 37 °C for 15 minutes.
  5. Place the stained cell solution on a slide, gently cover with a coverslip, and examine under a microscope.

Storage