Propidium Iodide (PI) Staining Solution
Research
Online Inquiry

Propidium Iodide (PI) Staining Solution

Cat.No: RESK-0526 Datasheet

Quantities:
- +
Product Details Related Products
Product Name Propidium Iodide (PI) Staining Solution
Catalog No. RESK-0526
Description Propidium iodide (PI) is a widely used red fluorescent nuclear stain. As an analog of ethidium bromide (EB), it intercalates between bases to bind DNA with little or no sequence preference, binding approximately one dye molecule per 4-5 DNA base pairs. PI also binds RNA, so nuclease (RNase) treatment is required to distinguish DNA from RNA staining. In aqueous solution, PI has excitation/emission maxima of 493/636 nm. Upon binding to nucleic acids, the fluorescence signal is enhanced 20-30 fold; the excitation maximum shifts ~30-40 nm toward the red and the emission maximum shifts ~15 nm toward the blue, giving excitation/emission maxima of 535/617 nm. Although PI has a relatively low molar absorptivity, its large Stokes shift allows simultaneous detection of nucleic acids and fluorophore-labeled antibodies when appropriate filters are used. PI is suitable for fluorescence microscopy, confocal microscopy, flow cytometry, and fluorometry. PI is membrane-impermeant and excluded from live cells, but it passes through compromised membranes to stain the nucleus. Based on this property, it is commonly combined with live-cell fluorescent probes such as Calcein-AM, Hoechst 33258, or Hoechst 33342 to simultaneously stain and identify live and dead cells in apoptosis-related research. PI can also serve as a counterstain in multiplex fluorescence staining and is compatible with a variety of cell-labeling techniques, including direct and indirect fluorescent antibody detection, mRNA in situ hybridization, cell-structure-specific fluorescent probes, and tissue staining. PI staining alone can also be used for cell cycle analysis. This product is a ready-to-dilute aqueous PI stock solution at 1 mg/mL; simply dilute with an appropriate buffer to obtain the working solution.
Intended Use Nuclear staining of dead or membrane-compromised cells; live/dead cell discrimination in apoptosis research (with live-cell probes); counterstaining in multiplex fluorescence labeling; cell cycle analysis by flow cytometry. For research use only.
Principle / Technology PI intercalates between DNA bases (about one dye per 4-5 base pairs, with little or no sequence preference). Because PI cannot cross intact cell membranes, it selectively stains dead or membrane-damaged cells. Nucleic-acid binding enhances fluorescence 20-30 fold and shifts Ex/Em from 493/636 nm to 535/617 nm.
Detection Method Fluorescence microscopy
Confocal microscopy
Flow cytometry (FL2 channel)
Fluorometry
Sample Type Cultured cells (live/dead discrimination), fixed cells, and tissue samples
Performance Range / Specifications English synonym: 2,7-Diamino-9-phenyl-10-(diethylaminopropyl)-phenanthridium iodide methiodide
CAS NO.: 25535-16-4
Formula: C27H34I2N4
Molecular weight: 668.4
Appearance: dark red crystals
Purity: ≥95% by HPLC
Solubility: soluble in water (1 mg/mL)
Fluorescence spectra: in aqueous solution, PI Ex/Em = 493/636 nm; PI-nucleic acid complex Ex/Em = 535/617 nm; fluorescence can be excited by a xenon lamp, a pump arc lamp, or a 488 nm argon-ion laser; typically detected in the FL2 channel of a flow cytometer
Specificity Stains nucleic acids with little or no sequence preference (one dye per 4-5 DNA base pairs); also binds RNA. Excluded from live cells with intact membranes; stains nuclei of dead or membrane-compromised cells.
Reaction Conditions / Protocol Dilute the 1 mg/mL stock solution with an appropriate buffer to the desired working concentration before use.
For DNA-specific staining, treat samples with RNase to remove RNA.
Samples fixed with formaldehyde, paraformaldehyde, or glutaraldehyde (cross-linking fixatives) require subsequent RNase treatment; samples fixed with methanol/acetic acid or acetone generally do not.
Components / Formulation Propidium iodide dissolved in water at 1 mg/mL.
Storage Conditions Store at 4°C, protected from light.
Shelf Life Stable for at least one year when stored at 4°C protected from light.
Package Specifications 1 mL
Product Form Aqueous solution (dark red)
Quality Control Purity ≥95% by HPLC.
Purity ≥95% by HPLC
Concentration 1 mg/mL in water
Molecular Weight 668.4
Shipping Conditions Shipped on wet ice.
Expiration Date / Stability Stable for at least one year at 4°C protected from light.
Regulatory / Compliance For research use only.
Compatibility Compatible with live-cell probes such as Calcein-AM, Hoechst 33258, and Hoechst 33342 for simultaneous live/dead cell staining.
Compatible with direct and indirect fluorescent antibody detection, mRNA in situ hybridization, cell-structure-specific fluorescent probes, and tissue staining.
Excitable by xenon lamp, pump arc lamp, or 488 nm argon-ion laser; detected in the FL2 channel of flow cytometers.
Recommended Buffer System Dilute with an appropriate aqueous buffer to the working concentration.
Application Notes / Precautions For red nuclear staining of dead cells, this PI staining solution or a 7-AAD staining solution is recommended.
PI is membrane-impermeant: it is excluded from live cells and stains only cells with compromised membranes; combine with live-cell probes for live/dead discrimination in apoptosis research.
PI also binds RNA; use RNase treatment when DNA-specific staining is required.
FAQ - Why does RNase treatment follow formaldehyde/paraformaldehyde/glutaraldehyde fixation but is usually unnecessary after methanol/acetic acid or acetone fixation? Cross-linking fixatives (formaldehyde, paraformaldehyde, glutaraldehyde) inactivate enzymes during cross-linking, so RNase must be added afterwards to remove RNA. Methanol/acetic acid or acetone fixation works by denaturation with only mild protein effects and allows re-solubilization after fixation, so RNase treatment is generally not required.
Protect from light.

For research use only, not for clinical use.

0
0

There is no product in your cart.