English
中文
CAS:28718-90-3
Appearance:Light yellow to green solid
Storage:Powder : -20℃, 2 years;In solvent(mother liquid): -20℃, 1 month; -80℃, 6 months
Purity:≥98%
Manual Download
It is A fluorescent dye that combines DNA sequences rich in A-T base pairs. DAPI is A fluorescent dye that binds to most of the A and T bases in DNA and is commonly used in fluorescence microscopy. Because DAPI can penetrate intact cell membranes, it can be used for staining both living and stationary cells. When DAPI is combined with double-stranded DNA, the maximum absorption wavelength is 358nm and the maximum emission wavelength is 461nm. The emission light of DAPI is blue, and the emission wavelengths of DAPI and green fluorescent protein GFP or Texas Red stain (red fluorescent stain) overlap only slightly, which can be used for multiple fluorescence staining on a single sample.
In general, the recommended DAPI working concentration for nuclear staining is 0.5-10μg/mL.
Note:
1)DAPI is irritating to the human body, please pay attention to proper protection.
2) Fluorescent dyes have the problem of quenching, it is recommended to complete the detection on the same day as possible after dyeing.
3) For your safety and health, please wear a lab coat and disposable gloves.
4) In general, DAPI is often used to fix cells or tissue sections for nucleation; Hoechst 33342 or Hoechst 33258 are recommended for the observation of the nuclei of living cells.
How to use (for reference only) :
1. Fixed cell or tissue staining: For fixed cell or tissue samples, after fixation, appropriate washing to remove the fixative. DAPI dyeing is usually done at the end of other dyeing. If no other coloring is required, DAPI is performed directly.
Dye it.
a) For adherent cells or tissue sections: add appropriate amount of DAPI staining solution and cover the sample.
For suspension cells: add at least 3 times the volume of the stain sample to be tested and mix well. Let stand at room temperature for 3-5 minutes.
b) Remove DAPI staining solution and wash with TBST, PBS or normal saline 2-3 times for 3-5 minutes each time.
c) Direct observation under fluorescence microscope or observation under fluorescence microscope after sealing. Excitation wavelength is 360nm, emission wavelength is 460nm.
2. Staining of living cells or tissues:
a) Add an appropriate amount of DAPI staining solution to the cell culture, about 1/10 of the volume of the cell medium, which must fully cover the sample to be stained. Usually, 1mL of dyeing solution should be added to a hole for a six-well plate, and 100μL dyeing solution should be added to a hole for a 96-well plate.
b) Cultured cells at 37℃ for 10 to 20 minutes.
c) Wash the cells twice with PBS or a suitable buffer.
d) Direct observation under fluorescence microscope or observation under fluorescence microscope after sealing. Excitation wavelength is 360nm, emission wavelength is 460nm.
