How to choose an appropriate TUNEL assay kit

2021-09-10Author:adminpraise:1

Commonly used TUNEL products on the market are mainly divided into two categories, one is fluorescent TUNEL products (suitable for fluorescence microscope or flow cytometer), and the other is HRP-DAB TUNEL assay kit (suitable for light microscope).

So, when choosing TUNEL products, we must check what kind of equipment we have. If a fluorescence microscope is available, choose fluorescent TUNEL products. If only light microscope is available, then to choose HRP-DAB TUNEL assay kit.

Compared with HRP-DAB TUNEL products, fluorescent TUNEL products have the following advantages:

● Shorter experiment time

● Easy to operate

● Easy to observe results

Elabscience® provide 5 kinds of fluorescent TUNEL kits, so how to choose the right TUNEL fluorescent kit?

Firstly, find out settings of fluorescence microscope, mainly the excitation and emission wavelength spectra.

The specific spectra information of each fluorescein is:

Fluorescein Excitation wavelength
(Ex)
Emission wavelength
(Em)
FITC 490 520
AF488 495 519
AF555 555 565
AF594 590 617
AF647 650 665
DAPI 372 456

DAPI is excited with ultraviolet light to emit blue light.

AF488 and FITC are excited with blue light to emit green light.

AF594 and AF555 are excited with green light to emit red light.

Secondly, find out whether the sample has auto-fluorescence.

According to the fluorescence of the experimental sample, the following principles should be followed when choosing the TUNEL kit.

a) If the experimental sample does not contain fluorescence, it is recommended to use One-step TUNEL Assay Kit (Green, FITC) or One-step TUNEL Assay Kit (Green, AF488).

b) If the experimental sample contain red fluorescence, it is recommended to use One-step TUNEL Assay Kit (Green, FITC) or One-step TUNEL Assay Kit (Green, AF488). Please note, the anti-quenching effect of AF488 is better than FITC. It is recommended to use One-step TUNEL Assay Kit (Green, AF488) if needing to observe the green fluorescence of the sample for a long time.

c) If the experimental sample contain green fluorescence, it is recommended to use One-step TUNEL Assay Kit (Red, AF594) or One-step TUNEL Assay Kit (Red, AF555).AF594 has less interference with GFP, if the sample has strong green fluorescence, One-step TUNEL Assay Kit (Red, AF594) is preferred.

d) If there are both red fluorescence and green fluorescence in the experimental sample, it is recommended to use One-step TUNEL Assay Kit (Red, AF647). However, AF647 has relatively high requirements for fluorescence microscopes, which should be confocal microscopes.