Pyronine is a Xanthene analog used as a dye and a bacterial and biological stain. It is a red band that appears during electrophoresis and is a cationic dye that intercalates RNA. It has also been demonstrated to build up in living cells' mitochondria. Pyronin Y is a fluorescent differential stain for nucleic acids that can be used on paraffin tissue sections in conjunction with Nancy-520 or Methyl Green.
Physicochemical Properties
| Molecular Formula | C17H19CLN2O |
| Molecular Weight | 302.7986 |
| Exact Mass | 302.118 |
| Elemental Analysis | C, 67.43; H, 6.32; Cl, 11.71; N, 9.25; O, 5.28 |
| CAS # | 92-32-0 |
| PubChem CID | 7085 |
| Appearance | Light brown to black solid powder |
| Melting Point | 250-260ºC(lit.) |
| Hydrogen Bond Donor Count | 0 |
| Hydrogen Bond Acceptor Count | 3 |
| Rotatable Bond Count | 1 |
| Heavy Atom Count | 21 |
| Complexity | 480 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | [Cl-].[O+]1C2C(=CC=C(N(C)C)C=2)C=C2C=1C=C(N(C)C)C=C2 |
| InChi Key | INCIMLINXXICKS-UHFFFAOYSA-M |
| InChi Code | InChI=1S/C17H19N2O.ClH/c1-18(2)14-7-5-12-9-13-6-8-15(19(3)4)11-17(13)20-16(12)10-14;/h5-11H,1-4H3;1H/q+1;/p-1 |
| Chemical Name | [6-(dimethylamino)xanthen-3-ylidene]-dimethylazanium;chloride |
| Synonyms | Pyronin Y; Pyronine |
| HS Tariff Code | 2934.99.03.00 |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| ln Vitro | In order to estimate formalin-fixed EL4 blank tumors, cellular enzyme-dispersed R3327-RNA content per cell in G-transporting adenocarcinoma cells, mouse splenocytes stimulated with concanavalin, and human peripheral blood stimulated with phytohemagglutinin, pyronin Y forms a fluorophore complex with double-stranded sequences, particularly RNA [1]. Give an explanation of the Jionin Y-based fluorescent staining technique. Human reticulocytes have been treated with this method to dye their RNA in preparation for sorting and process analysis [2]. This dye gathers in the mitochondria of live cells. Jiaoning Y forms additional mitochondria in the mitochondria of cultivated cells when it is almost entirely localized there at 1.7 to 3.3 μM! Rhodamine 123 probe. PY is not harmful at this dose, but it will stop cell division and induce apoptosis [3]. For a considerable amount of time, paraffin tissue sections have been differentially stained using pyronin Y in conjunction with other dyes like methyl green. Cell nuclei appeared non-fluorescent in methyl green-pyronin Y stained sections, whereas red blood cells, elastic fibers, amygdala acinar cell zymogen granules, liver cell surface membranes, and renal tubular cells all displayed intense green and/or red fluorescence[4]. Double-stranded RNA (dsRNA) such as messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA) are bound by pyrin Y [5]. |
| References |
[1]. Flow cytometric analysis of RNA content in different cell populations using pyronin Yand methyl green. Cytometry. 1982 Jul;3(1):28-35. [2]. Flow cytometry of human reticulocytes based on RNA fluorescence. Cytometry. 1981 Mar;1(5):313-20. [3]. 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]. Pyronin Y as a fluorescent stain for paraffin sections. Histochem J. 2002 Jun-Jul;34(6-7):299-303. [5]. Spectral phasor analysis of Pyronin Y labeled RNA microenvironments in living cells. Biomed Opt Express. 2013 Jan 1;4(1):171-7. |
| Additional Infomation |
Pyronin Y is an organic chloride salt having 6-(dimethylamino)-N,N-dimethyl-3H-xanthen-3-iminium as the cation. Used with methyl green to selectively demonstrate RNA (red) in contrast to DNA (green) with the Unna-Pappenheim method. It has a role as a histological dye. It is an iminium salt and an organic chloride salt. It contains a pyronin Y cation. Xanthene dye used as a bacterial and biological stain. Synonyms: Pyronin; Pyronine G; Pyronine Y. Use also for Pyronine B. which is diethyl-rather than dimethylamino-. |
Solubility Data
| Solubility (In Vitro) |
DMSO : ~25 mg/mL (~82.56 mM) H2O : ~4 mg/mL (~13.21 mM) |
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 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 | 6.6050 mL | |
| 10 mM | 0.3303 mL | 1.6513 mL | 3.3025 mL |