Pyronin Y (Pyronine G) (Synonyms: C.I. 45005) |
| Katalog-Nr.GC30149 |
Pyronin Y (Pyronin G) (Pyronin G) ist ein kationischer Farbstoff, der RNA interkaliert und verwendet wurde, um auf Zellstrukturen einschließlich RNA, DNA und Organellen abzuzielen.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 92-32-0
Sample solution is provided at 25 µL, 10mM.
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Pyronin Y is an intercalating cationic dye that shows specificity towards RNA.
Pyronin Y forms fluorescent complexes with double-stranded nucleic acids, especially RNA, enabling semi-quantitative analysis of cellular RNA in flow cytometry, to estimate the RNA content per cell in formalin fixed EL4 leukosis tumor cells, enzyme dispersed R3327-G rat prostatic adenocarcinoma cells, mouse spleen cells stimulated with concanavalin A, and human peripheral blood lymphocytes stimulated with phytohemagglutinin[1]. A fluorescent staining procedure based on pyronin Y is described. The technique has been used to stain RNA in human reticulocytes for subsequent flow analysis and sorting[2]. In viable cells this dye also accumulates in mitochondria. At a concentration of 1.7 to 3.3 μM, pyronin Y is localized almost exclusively in mitochondria of cultured cells, similar to another mitochondria! probe, rhodamine 123. At that concentration PY is not toxic but suppressed cell growth, arresting cells[3]. Pyronin Y has long been used, in combination with other dyes such as Methyl Green, as a differential stain for nucleic acids in paraffin tissue sections. In sections stained with Methyl Green-pyronin Y, red blood cells, elastic fibre of blood vessels, zymogen granules of pancreatic acinar cells, surface membrane of heptocytes and kidney tubular cells showed strikingly strong green and/or red fluorescence, while the nuclei of cells appeared non-fluorescent[4].
[1]. Pollack A, et al. Flow cytometric analysis of RNA content in different cell populations using pyronin Yand methyl green. Cytometry. 1982 Jul;3(1):28-35. [2]. Tanke HJ, et al. Flow cytometry of human reticulocytes based on RNA fluorescence. Cytometry. 1981 Mar;1(5):313-20. [3]. Darzynkiewicz Z, et al. Cytostatic and cytotoxic properties of pyronin Y: relation to mitochondrial localization of the dye and its interaction with RNA. Cancer Res. 1986 Nov;46(11):5760-6. [4]. Li B, et al. Pyronin Y as a fluorescent stain for paraffin sections. Histochem J. 2002 Jun-Jul;34(6-7):299-303.
This plan only provides a guide, please modify it to meet your specific needs.
1. Prepare Pyronin Y staining solution
(1) Prepare Pyronin Y dye stock solution: Use DMSO/sterile water to dissolve the solid powder and prepare a 1-5mM Pyronin Y dye stock solution.
Note: The stock solution prepared with sterile water needs to be filtered with a 0.22μm filter membrane. It is recommended to aliquot Pyronin Y stock solution and store it in the dark at -20°C to avoid repeated freezing and thawing.
(2) Working solution preparation: Dilute the stock solution with a suitable buffer solution (such as HBSS or PBS) and prepare a concentration of 1-5μM Pyronin Y working fluid.
Note: Please adjust the concentration of Pyronin Y working fluid according to the actual situation and prepare it for use.
2. Cell staining
2.1 Suspension cells (taking 6-well plate as an example)
(1) Centrifuge suspended cells at 1000g for 3-5 minutes. Discard the supernatant and wash twice with PBS for 5 minutes each time.
(2) Add 1 mL of Pyronin Y dye working solution and incubate at room temperature in the dark for 1-5 minutes. Different cells have different optimal culture times.
(3) After the incubation, centrifuge at 1000g for 5 minutes, remove the supernatant, add PBS and wash 2-3 times, 5 minutes each time.
(4) Use serum-free cell culture medium or PBS to resuspend the cells and observe them through fluorescence microscopy or flow cytometry.
2.2 Adherent cells
(1) Culture adherent cells on sterile coverslips.
(2) Remove the coverslip from the culture medium, suck out the excess culture medium, and place the coverslip in a humid environment.
(3) Add 100μL of Pyronin Y dye working solution from one corner of the coverslip, shake gently to evenly cover all cells with the dye, and incubate at room temperature in the dark for 1-5 minutes.
(4) Aspirate away the dye working solution, wash the coverslip 2 to 3 times with culture solution, and observe with a fluorescence microscope.
3. Microscope detection: The maximum excitation light of Pyronin Y is 540-550nm, and the maximum emission light is 560-580nm.
Notes:
① Pyronin Y is suitable for staining paraffin sections. It is recommended to prepare 5% Pyronin Y staining solution in PBS and incubate at room temperature in dark for 5-10 minutes;
② Fluorescent dyes all have quenching problems. Please try to avoid light as much as possible to slow down fluorescence quenching;
③ For your safety and health, please wear laboratory clothes and disposable gloves when operating.
References:
[1]. Taro Mannen, Tetsuro Hirose. RNase Sensitivity Screening for Nuclear Bodies with RNA Scaffolds in Mammalian Cells. 2017 Apr 20;7(8):e2232. doi: 10.21769/BioProtoc.2232.
[2]. Bo Li, Ying Wu, Xiao-Ming Gao. Pyronin Y as a fluorescent stain for paraffin sections. 2002 Jun-Jul;34(6-7):299-303. doi: 10.1023/a:1023325213198.
| Cas No. | 92-32-0 | SDF | |
| Überlieferungen | C.I. 45005 | ||
| Canonical SMILES | CN(C1=CC2=[O+]C3=C(C=CC(N(C)C)=C3)C=C2C=C1)C.[Cl-] | ||
| Formula | C17H19ClN2O | M.Wt | 302.8 |
| Löslichkeit | DMSO : 25 mg/mL (82.56 mM);Water : 4 mg/mL (13.21 mM) | 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 | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 3.3025 mL | 16.5125 mL | 33.0251 mL |
| 5 mM | 0.6605 mL | 3.3025 mL | 6.605 mL |
| 10 mM | 0.3303 mL | 1.6513 mL | 3.3025 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 ddH2O, 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: >50.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 36 reference(s) in Google Scholar.)
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