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Journal of Histochemistry and Cytochemistry, Vol. 50, 1421-1424, October 2002, Copyright © 2002, The Histochemical Society, Inc.

BRIEF REPORT

Effects of Different Fixatives on ß-Galactosidase Activity

Wenbin Maa, Keith Rogersb, Berton Zbara, and Laura Schmidtc
a Laboratory of Immunobiology, SAIC–Frederick, Inc., NCI-Frederick, Frederick, Maryland
b Pathology and Histology Laboratory, SAIC–Frederick, Inc., NCI-Frederick, Frederick, Maryland
c Intramural Research Support Program, SAIC–Frederick, Inc., NCI-Frederick, Frederick, Maryland

Correspondence to: Laura Schmidt, IRSP, SAIC–Frederick, NCI–Frederick, Building 560, Room 12-69, Frederick, MD 21702. E-mail: schmidtl@mail.ncifcrf.gov

ß-Galactosidase (ß-Gal) staining is widely used to demonstrate specific gene expression during evaluation of gene targets in vivo. This technique is extremely sensitive to fixation. Optimal fixation conditions are necessary to obtain the maximal ß-Gal activity. In this experiment, Carnoy's and three different aldehyde fixatives were used at different temperatures and over different time points. Kidneys from LacZ-stop-human alkaline phosphatase (ZA/P) double reporter mice were used to generate positive material for the experiment. The results show that glutaraldehyde combinative solution (LacZ) produced the most consistent and reliable results. Paraformaldehyde and formaldehyde were effective as fixatives only at 4C for a period of less than 4 hr, and Carnoy's solution destroyed ß-Gal activity. (J Histochem Cytochem 50:1421–1424, 2002)

Key Words: ß-galactosidase, X-gal staining, fixative, temperature


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