How to choose an appropriate TUNEL assay kit

2021-09-10Author:adminpraise:3

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

Elab Fluor® 488

495 519
Elab Fluor® 555 555 565
Elab Fluor® 594 590 617
Elab Fluor® 647 650 665
DAPI 372 456

DAPI is excited with ultraviolet light to emit blue light.

Elab Fluor®488 and FITC are excited with blue light to emit green light.

Elab Fluor®594 and Elab Fluor®555 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, Elab Fluor®488).

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, Elab Fluor®488). Please note, the anti-quenching effect of Elab Fluor®488 is better than FITC. It is recommended to use One-step TUNEL Assay Kit (Green, Elab Fluor®488) 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, Elab Fluor®594) or One-step TUNEL Assay Kit (Red, Elab Fluor®555).Elab Fluor®594 has less interference with GFP, if the sample has strong green fluorescence, One-step TUNEL Assay Kit (Red, Elab Fluor®594) 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, Elab Fluor®647). However, Elab Fluor®647 has relatively high requirements for fluorescence microscopes, which should be confocal microscopes.