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CAS:23491-45-4
Appearance:Light yellow to green solid
Storage:Powder : -20℃, 2 years;In solvent(mother liquid): -20℃, 1 month; -80℃, 6 months
Purity:HPLC≥98%
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Hoechst 33258 is a blue fluorescent dye that can penetrate cell membranes, which emits strong blue fluorescence upon embedding in double-stranded DNA and is less toxic to cells. Hoechst 33258 is commonly used to detect apoptosis, observed by fluorescence microscopy or flow cytometry after staining, and can also be used for ordinary nuclear staining, or conventional DNA staining. Hoechst 33258 staining is often used to detect apoptosis, which is observed by fluorescence microscopy or flow cytometry after staining. Hoechst 33258 is also commonly used for conventional nuclear staining, or conventional DNA staining. Hoechst 33258 has a maximum excitation wavelength of 346nm and a maximum emission wavelength of 460nm. When combined with double-stranded DNA, Hoechst 33258 has a maximum excitation wavelength of 352nm and a maximum emission wavelength of 461nm.
Instructions (for reference only) :
1. For fixed cells or tissues:
a. For cell or tissue samples, after fixation, appropriate washing to remove the fixative. Subsequently, if immunofluorescence staining is required, immunofluorescence staining should be performed first, and Hoechst 33258 staining should be performed according to the following steps after staining. If no additional staining is required, the subsequent staining with Hoechst 33258 is performed directly.
b. For adherent cells or tissue sections, add a small amount of Hoechst 33258 working solution to cover the sample; For suspension cells, add at least 3 times the volume of the sample to be dyed and mix well. Let stand at room temperature for 3-5 minutes.
c. Remove Hoechst 33258 staining solution and wash with TBST, PBS or normal saline 2-3 times for 3-5 minutes each time.
d. Direct observation under fluorescence microscope or observation under fluorescence microscope after sealing. When apoptosis occurs, the nuclei of apoptotic cells are dense and densely stained, or fragmented and densely stained.
2. For living cells or tissues:
a. Add the appropriate amount of Hoechst 33258 working liquid, must fully cover the sample to be dyed, usually add 1mL of working liquid for each hole of the six-well plate, and add 100μL of working liquid for each hole of the 96-well plate.
b. Culture at a temperature suitable for cell culture for 20-30 minutes. Discard the dye solution and wash it with PBS or culture solution 2-3 times for fluorescence detection.
Note:
All fluorescent dyes have the problem of quenching. In order to slow down the quenching of fluorescence, anti-fluorescence attenuation tablets can be used. It is recommended to complete the test on the same day as possible after staining, and live cells or tissues should be observed immediately after staining.
For your safety and health, please wear a lab coat and disposable gloves.以上翻译结果来自有道神经网络翻译（YNMT）· 通用场景
