Description
The Prodidium Iodide (PI) Staining Solution may be used to assess plasma membrane (PM) integrity in Annexin V apoptosis assays. PI is a fluorescent vital dye that stains DNA. It does not cross the PM of cells that are viable or in the early stages of apoptosis because they maintain PM integrity. In contrast those cells in the late stages of apoptosis or already dead have lost PM integrity and are permeable to PI. PI is detected in the orange range of the spectrum using a 562-588 nm band pass filter. Annexin V binds to cells early in apoptosis, and continues to be bound through cell death. PI is used in two-color Annexin V flow cytometric assays to distinguish cells that are in the earlier stages of apoptosis (Annexin V positive, PI negative) from those that are in the later stages of apoptosis or already dead (Annexin V positive, PI positive).
Suggested Companion Products
Purified NA/LE Mouse Anti-Human CD95 DX2RUO 0.5mgCat No: 555670
FITC Streptavidin RUO 0.5mgCat No: 554060
FITC Annexin V RUO 200TestsCat No: 556419
Biotin Annexin V RUO 200TestsCat No: 556417
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Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light.Do not freeze.This preparation contains no preservatives, thus it should be handled under aseptic conditions.The PI Staining Solution is composed of 50 µg PI/ml in PBS (pH 7.4) and is 0.2 µm sterile filtered.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- This reagent has been pre-diluted for use at the recommended Volume per Test.We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
Recommended Assay Procedure:
The PI Staining Solution is designed for use in two-color Annexin V flow cytometric assays. It can be used in conjunction with Annexin V conjugated to fluorescein isothiocyanate (Annexin V-FITC, Cat. No. 556419) or biotinylated Annexin V (Annexin V-Biotin, Cat. No. 556417). Suggested amounts to use are 10 µl/test (1x10e6 cells).
Caution:Propidium Iodide is a potential carcinogen. Avoid contact with skin and eyes. Wear suitable protective clothing, gloves, and eye/face protection.
