Propidium iodide (Synonyms: PI) |
رقم الكتالوجGC14228
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يعد اليوديد بروبيديوم (PI) جزيء أحمر صغير يترابط مع الحمض النووي ولكنه لا يستطيع اختراق الخلايا التى تحتفظ بغشاء خلوى سليم.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 25535-16-4
Sample solution is provided at 25 µL, 10mM.
Propidium iodide (PI) is a small red-fluorescent molecule that binds to DNA but cannot passively traverse into cells that possess an intact plasma membrane. Propidium iodide uptake versus exclusion can be used to discriminate dead cells. PI is excited by wavelengths between 400 and 600 nm and emits light between 600 and 700 nm [1]. PI intercalates to DNA with no sequence preference with one dye molecule per four to five base pairs [2]. When bound to DNA fluorescence of PI is enhanced 20- to 30-fold [3]. PI binds stoichiometrically to nucleic acids so that fluorescence emission is proportional to the DNA (and RNA, which has to be removed if DNA is to be measured) content of a cell [4].
References:
[1]. Crowley L C, Scott A P, Marfell B J, et al. Measuring cell death by propidium iodide uptake and flow cytometry[J]. Cold Spring Harbor Protocols, 2016, 2016(7): pdb. prot087163.
[2]. Waring M J. Complex formation between ethidium bromide and nucleic acids[J]. Journal of molecular biology, 1965, 13(1): 269-282.
[3]. Arndt-Jovin D J, Jovin T M. Fluorescence labeling and microscopy of DNA[J]. Methods in cell biology, 1989, 30: 417-448.
[4]. Riccardi C, Nicoletti I. Analysis of apoptosis by propidium iodide staining and flow cytometry[J]. Nature protocols, 2006, 1(3): 1458-1461.
Procedure for Propidium iodide staining [1]
Fluorochrome solution: 0.1% sodium citrate (wt/v), 0.1% Triton X-100 (v/v), 50 mg l−1 Propidium iodide in deionized/distilled water). Fluorochrome solution can be kept in the dark at 4 °C for months.
DNA staining solution: Dissolve 200 μg of Propidium iodide in 10 ml of PBS. Add 2 mg of DNase free RNase.
CRITICAL：Prepare fresh staining solution just before use.
- Suspend cells at 1–2 × 106 cells ml−1 in 1 ml of PBS in 12×75 tubes.
- Centrifuge at 200g for 5 min at room temperature.
- Aspirate off the PBS.
- Follow the quick method for thymocytes and non-adherent mononuclear cells (option A) and the standard method for multinuclear cells growing in suspension and for adherent cells (option B).
- Quick method (direct DNA staining in PI hypotonic solution)
i) Gently resuspend the cell pellet in 1 ml of fluorochrome solution.
ii) Place the tubes in the dark at 4 °C, before flow cytometry analysis, for at least 1 h and no longer than 24 h.
Note: One hour is necessary for appropriate staining of the nuclei; cells can be maintained for 24 h, in the dark at 4 °C without any substantial change in DNA profile.
B. Standard method (PI staining after alcoholic fixation)
i) Resuspend cell pellet in 500 μl of PBS.
ii) Fix cells by adding 4.5 ml of 70% (v/v) cold ethanol to the cell suspension keeping the tubes on ice.
PAUSE POINT: Cells can be stored in ethanol solution at −20 °C for several weeks.
iii) Centrifuge at 400g for 5 min and remove the supernatant (ethanol solution).
iv) Wash cells in 5 ml of PBS and centrifuge at 400g for 5 min.
CRITICAL STEP: Cells with extensive DNA degradation can be directly resuspended in DNA staining solution without any further treatment.
v) If DNA is not extensively degraded, resuspend cells in 0.5 ml of PBS and add 0.5 ml of DNA extraction buffer. Incubate at room temperature for 5 min and centrifuge at 400g for 5 min.
vi） Remove the supernatant and resuspend cells in 1 ml of DNA staining solution.
vii） Incubate resuspended cells for at least 30 min at room temperature in the dark.
- Analyze cells by flow cytometry. Use 488-nm laser line for excitation. Measure red fluorescence (>600 nm) and side scatter. Collect at least 20,000 events. Gate-out residual debris. Measure hypodiploid and diploid DNA peaks.
This protocol only provides a guideline, and should be modified according to your specific needs
[1]. Riccardi C, Nicoletti I. Analysis of apoptosis by propidium iodide staining and flow cytometry[J]. Nature protocols, 2006, 1(3): 1458-1461.
| Cas No. | 25535-16-4 | SDF | |
| المرادفات | PI | ||
| Chemical Name | 3,8-diamino-5-(3-(diethyl(methyl)ammonio)propyl)-6-phenylphenanthridin-5-ium iodide | ||
| Canonical SMILES | NC1=CC2=[N+](C(C3=CC=CC=C3)=C(C=C4N)C(C=C4)=C2C=C1)CCC[N+](CC)(C)CC.[I-].[I-] | ||
| Formula | C27H34I2N4 | M.Wt | 668.39 |
| الذوبان | 100 mg/mL in DMSO (Need ultrasonic); 5mg/mL in Water (Need ultrasonic). | Storage | Store at -20°C,protect from light |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.4961 mL | 7.4807 mL | 14.9613 mL |
| 5 mM | 299.2 μL | 1.4961 mL | 2.9923 mL |
| 10 mM | 149.6 μL | 748.1 μL | 1.4961 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
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Average Rating: 5 (Based on Reviews and 21 reference(s) in Google Scholar.)