Antifade Mounting Medium (Containing Hoechst 33258 and PI) (C008B-248812)

Hoechst 33258 is also known as bisBenzimide H 33258 or HOE 33258, with the molecular formula C₂₅H₂₄N₆O·3HCl, a molecular weight of 533.88 and CAS Number 23491-45-4. It is a blue fluorescent dye that can penetrate cell membranes with low cytotoxicity, and is commonly used for the detection of cell apoptosis. After staining, observations can be made under a fluorescence microscope or detections via a flow cytometer. Hoechst 33258 is also applicable for general nuclear staining and DNA staining. The maximum excitation wavelength of Hoechst 33258 is 346 nm and its maximum emission wavelength is 460 nm; upon binding to double-stranded DNA, its maximum excitation wavelength becomes 352 nm and the maximum emission wavelength 461 nm.Propidium Iodide, abbreviated as PI, is a nucleic acid staining reagent capable of staining DNA and RNA. As an analog of ethidium bromide, it intercalates into double-stranded DNA or RNA to form PI-DNA or PI-RNA complexes, which emit red fluorescence. Propidium Iodide (PI) is a dark red solid powder with the molecular formula C₂₇H₃₄I₂N₄, a molecular weight of 668.4 and CAS Number 25535-16-4. The excitation and emission wavelengths of the PI-DNA complex are 535 nm and 615 nm, respectively.Excitation and Emission Spectra of Hoechst 33258-DNA Complex (Left) and PI-DNA Complex (Right)Although PI cannot penetrate the membranes of live cells, it can cross damaged cell membranes to stain cell nuclei and other structures, and is therefore often used in combination with Hoechst 33258 for the detection of cell apoptosis and necrosis. PI can also be used for nuclear staining following routine cell permeabilization.Antifade Mounting Medium (Containing Hoechst 33258 and PI) is a mounting reagent containing Hoechst 33258, a blue fluorescent nuclear dye, and PI, a red fluorescent dye, with the ability to mitigate fluorescent quenching. This mounting medium not only reduces the fluorescent quenching of various common fluorescent dyes, but also enables simultaneous Hoechst 33258 and PI staining during the mounting process. This reagent is for research use only and not intended for clinical diagnosis or any other applications.Materials to Be Prepared by the User:1. Fluorescence microscope, micropipette2. Distilled water, PBS or normal salineOperating Procedures (For Reference Only):1. Adherent cell samples:a. Aspirate all liquid completely after staining.b. Add one drop of this reagent onto a glass slide, cover with a coverslip with adherent cells attached to make the cells contact the mounting medium, and avoid bubbles as much as possible.c. The cell samples can then be observed directly under a fluorescence microscope.2. Tissue sections:a. Aspirate all liquid completely after staining.b. Add one drop of this reagent onto the tissue section, cover with a coverslip to make the section contact the mounting medium, and avoid bubbles as much as possible.c. The tissue sections can then be observed directly under a fluorescence microscope.3. Other samples:Refer to the procedures for cell samples or tissue sections for the operation of other samples.Staining Results:In apoptotic cells, the cell nuclei appear densely stained or show a densely stained fragmented pattern.Precautions:1. All fluorescent substances are prone to quenching; stained samples should be stored in the dark.2. For optimal staining and identification results, it is recommended to complete the detection within 20–30 minutes after staining.3. The use of anti-fade mounting medium can mitigate quenching, yet storage in the dark is still highly recommended.4. This product is intended exclusively for scientific research by professional personnel, and shall not be used for clinical diagnosis or treatment, food or pharmaceutical applications, nor stored in ordinary residential premises.5. For your safety and health, wear a lab coat and disposable gloves during operation.

Specifications & Purity: Hoechst 33258 is also known as bisBenzimide H 33258 or HOE 33258, with the molecular formula C₂₅H₂₄N₆O·3HCl, a molecular weight of 533.88 and CAS Number 23491-45-4. It is a blue fluorescent dye that can penetrate cell membranes with low cytotoxicity, and is commonly used for the detection of cell apoptosis. After staining, observations can be made under a fluorescence microscope or detections via a flow cytometer. Hoechst 33258 is also applicable for general nuclear staining and DNA staining. The maximum excitation wavelength of Hoechst 33258 is 346 nm and its maximum emission wavelength is 460 nm; upon binding to double-stranded DNA, its maximum excitation wavelength becomes 352 nm and the maximum emission wavelength 461 nm.Propidium Iodide, abbreviated as PI, is a nucleic acid staining reagent capable of staining DNA and RNA. As an analog of ethidium bromide, it intercalates into double-stranded DNA or RNA to form PI-DNA or PI-RNA complexes, which emit red fluorescence. Propidium Iodide (PI) is a dark red solid powder with the molecular formula C₂₇H₃₄I₂N₄, a molecular weight of 668.4 and CAS Number 25535-16-4. The excitation and emission wavelengths of the PI-DNA complex are 535 nm and 615 nm, respectively.Excitation and Emission Spectra of Hoechst 33258-DNA Complex (Left) and PI-DNA Complex (Right)Although PI cannot penetrate the membranes of live cells, it can cross damaged cell membranes to stain cell nuclei and other structures, and is therefore often used in combination with Hoechst 33258 for the detection of cell apoptosis and necrosis. PI can also be used for nuclear staining following routine cell permeabilization.Antifade Mounting Medium (Containing Hoechst 33258 and PI) is a mounting reagent containing Hoechst 33258, a blue fluorescent nuclear dye, and PI, a red fluorescent dye, with the ability to mitigate fluorescent quenching. This mounting medium not only reduces the fluorescent quenching of various common fluorescent dyes, but also enables simultaneous Hoechst 33258 and PI staining during the mounting process. This reagent is for research use only and not intended for clinical diagnosis or any other applications.Materials to Be Prepared by the User:1. Fluorescence microscope, micropipette2. Distilled water, PBS or normal salineOperating Procedures (For Reference Only):1. Adherent cell samples:a. Aspirate all liquid completely after staining.b. Add one drop of this reagent onto a glass slide, cover with a coverslip with adherent cells attached to make the cells contact the mounting medium, and avoid bubbles as much as possible.c. The cell samples can then be observed directly under a fluorescence microscope.2. Tissue sections:a. Aspirate all liquid completely after staining.b. Add one drop of this reagent onto the tissue section, cover with a coverslip to make the section contact the mounting medium, and avoid bubbles as much as possible.c. The tissue sections can then be observed directly under a fluorescence microscope.3. Other samples:Refer to the procedures for cell samples or tissue sections for the operation of other samples.Staining Results:In apoptotic cells, the cell nuclei appear densely stained or show a densely stained fragmented pattern.Precautions:1. All fluorescent substances are prone to quenching; stained samples should be stored in the dark.2. For optimal staining and identification results, it is recommended to complete the detection within 20–30 minutes after staining.3. The use of anti-fade mounting medium can mitigate quenching, yet storage in the dark is still highly recommended.4. This product is intended exclusively for scientific research by professional personnel, and shall not be used for clinical diagnosis or treatment, food or pharmaceutical applications, nor stored in ordinary residential premises.5. For your safety and health, wear a lab coat and disposable gloves during operation.

Grade: BioReagent;Biological Stain;for Microscopy

Storage Temperature: Protected from light;Store at -20°C

Shipped in: Ice chest + Ice pads

UPC:

51284123

Weight:

1.23 Ounces

HazmatClass:

No

Quantity:

1

Unit of Measurement:

EA

WeightUOM:

LB

MPN:

A1511203-5ml

Country of Origin:

China

Product Size:

5ml

Harmonized Tariff Code:

2931.90.90