Effects of different fixatives on β-galactosidase activity

被引:29
作者
Ma, WB
Rogers, K
Zbar, B
Schmidt, L
机构
[1] NCI Frederick, SAIC Frederick, IRSP, Ft Detrick, MD 21702 USA
[2] NCI Frederick, SAIC Frederick, Immunobiol Lab, Ft Detrick, MD 21702 USA
[3] NCI Frederick, SAIC Frederick, Pathol & Histol Lab, Ft Detrick, MD 21702 USA
关键词
beta-galactosidase; X-gal staining; fixative; temperature;
D O I
10.1177/002215540205001015
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
beta-Galactosidase (beta-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 P-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 P-Gal activity.
引用
收藏
页码:1421 / 1424
页数:4
相关论文
共 7 条
[1]  
Chai Y, 2000, DEVELOPMENT, V127, P1671
[2]   Z/AP, a double reporter for Cre-mediated recombination [J].
Lobe, CG ;
Koop, KE ;
Kreppner, W ;
Lomeli, H ;
Gertsenstein, M ;
Nagy, A .
DEVELOPMENTAL BIOLOGY, 1999, 208 (02) :281-292
[3]   Murine model of autosomal dominant retinitis pigmentosa generated by targeted deletion at codon 307 of the rds-peripherin gene [J].
McNally, N ;
Kenna, PF ;
Rancourt, D ;
Ahmed, T ;
Stitt, A ;
Colledge, WH ;
Lloyd, DG ;
Palfi, A ;
O'Neill, B ;
Humphries, MM ;
Humphries, P ;
Farrar, GJ .
HUMAN MOLECULAR GENETICS, 2002, 11 (09) :1005-1016
[4]   Stage-and tissue-specific expression of a Col2a1-Cre fusion gene in transgenic mice [J].
Sakai, K ;
Hiripi, L ;
Glumoff, V ;
Brandau, O ;
Eerola, R ;
Vuorio, E ;
Bösze, Z ;
Fässler, R ;
Aszódi, A .
MATRIX BIOLOGY, 2001, 19 (08) :761-767
[5]  
Sheehan DC., 1980, Theory and Practice of Histotechnology
[6]   Generalized lacZ expression with the ROSA26 Cre reporter strain [J].
Soriano, P .
NATURE GENETICS, 1999, 21 (01) :70-71
[7]   A highly efficient ligand-regulated Cre recombinase mouse line shows that LoxP recombination is position dependent [J].
Vooijs, M ;
Jonkers, J ;
Berns, A .
EMBO REPORTS, 2001, 2 (04) :292-297