Home
Resources
Publications
A Fluorimetric Method Using Fluorescein di-beta-D-galactopyranoside for Quantifying the Senescence-Associated Beta-Galactosidase Activity in Human Foreskin Fibroblast Hs68 Cells

A Fluorimetric Method Using Fluorescein di-beta-D-galactopyranoside for Quantifying the Senescence-Associated Beta-Galactosidase Activity in Human Foreskin Fibroblast Hs68 Cells

Nae-Cherng Yang, Miao-Lin Hu

Anal Biochem. 2004 Feb 15;325(2):337-43.

PMID: 14751269

Abstract:

The senescence-associated beta-galactosidase (SA-betaG) assay is one of the few accepted markers of cell aging. However, the cytochemical method using 5-bromo-4-chloro-3-indolyl beta-D-galactopyranoside (X-Gal) as substrate is limited in sensitivity and is only semiquantitative. Here, we modified the X-Gal method by replacing X-Gal with fluorescein di-beta-D-galactopyranoside (FDG) as substrate for SA-betaG, and the activity was measured fluorimetrically. We showed in Hs68 cells that the FDG fluorescein fluorescence increased with increasing passages of the cells in parallel with the X-Gal method. A major advantage of the FDG method is that it is a quantitative method for the SA-betaG activity. For example, we showed that the FDG fluorescein in p30(+1) of Hs68 cells was generally stronger than that in p26(+1) cells, whereas the X-Gal method gave similar results (95 and 100%) for p26(+1) and p30(+1) cells. The FDG method was precise with a relative standard deviation lower than 10%. We further demonstrated that FDG and X-Gal could be added simultaneously for SA-betaG assay because the FDG fluorescein diffused readily through formaldehyde-fixed cell membrane and could be detected in the suspension buffer. Thus, a double-substrate method, i.e., X-Gal for rapid qualitative assay and FDG for quantitative assay, can be conducted simultaneously to provide a simple and reliable assay of SA-betaG activity as a marker of cell aging.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP17817208	Fluorescein di(β-D-galactopyranoside)		17817-20-8	Price

As a specialist in analytical chemical Products, Alfa Chemistry offers consumers a wealth of raw materials and a full range of technologies and services.

QUICK LINKS

HEADQUARTERS

Tel:

Fax:

Email: