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CAS:875756-97-1
Appearance:Solid
Storage:Powder : -20℃, 1 year;In solvent(mother liquid): -20℃, 1 month; -80℃, 6 months
Purity:≥98%
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Is a blue fluorescent dye that can penetrate cell membranes, it emits strong blue fluorescence after embedding double-stranded DNA, and is less toxic to cells. Hoechst 33342 staining is often used to detect apoptosis, which is observed by fluorescence microscopy or flow cytometry after staining. Hoechst 33342 is also commonly used for conventional nuclear staining, or conventional DNA staining. Hoechst 33342 has a maximum excitation wavelength of 346nm and a maximum emission wavelength of 460nm. Hoechst 33342 combined with double-stranded DNA has a maximum excitation wavelength of 350nm and a maximum emission wavelength of 461nm.
Instructions for use (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 33342 staining should be performed according to the following steps after staining. If no additional staining is required, the subsequent Hoechst 33342 staining is performed directly.
b. For adherent cells or tissue sections, add a small amount of Hoechst 33342 staining 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 33342 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 33342 staining solution, which must fully cover the sample to be dyed, usually add 1mL dyeing solution for each hole of the six-well plate and 100 microliters dyeing solution 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: Fluorescent dyes have the problem of quenching, in order to slow down the fluorescence quenching can be used anti-fluorescence attenuation tablets. 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.
Examples of using this product（for reference only）
In Vitro:
Cell（LUAD cell， 10 ug/ml Hochest 33342， RT， 20min）:
Hochest 33342 (Solarbio, Beijing, China) staining was performed to evaluate the apoptosis of LUAD cell induced by onvansertib treatment. Briefly, cells were cultured in in 6-well plates, stained with 10 ug/ml Hochest 33342 at room temperature for 20 minutes, washed with PBS, and observed under EVOS M7000 fluorescence microscope.
Reference：
Wang R, Hou Y, Geng G, Zhu X, Wang Z, Cai W, Ye J, Zhao S, Mi Y, Jiang J. Onvansertib inhibits the proliferation and improves the cisplatin-resistance of lung adenocarcinoma via β-catenin/c-Myc signaling pathway. Am J Cancer Res. 2023 Feb 15;13(2):623-637. PMID: 36895968; PMCID: PMC9989612.
