Y. Tsuji et al., RETINOBLASTOMA SUSCEPTIBILITY GENE-PRODUCT RETRIEVAL IN FORMALIN-FIXED, PARAFFIN-EMBEDDED TISSUE - A HEATING METHOD FOR ENHANCING IMMUNOHISTOCHEMICAL STAINING, Acta histochemica et cytochemica, 31(3), 1998, pp. 193-196
We examined immunohistochemical methods to visualize the retinoblastom
a susceptibility gene product (RB) in formalin-fixed, paraffin-embedde
d sections of the U2-OS and Saos-2 human osteosarcoma cell lines. Immu
nocytochemical studies showed clear staining with an anti-RB antibody
in 88% of the smeared U2-OS cells and in none of the smeared Saos-2 ce
lls. Antigen retrieval methods evaluated included microwave-oven and a
utoclave pretreatment of paraffin-embedded sections immersed in Dulbec
co's phosphate buffered saline (pH 7.4) prior to incubation with the p
rimary antibody. Strong and discrete staining of U2-OS cell nuclei for
RB was seen after autoclave treatment, stronger than that seen follow
ing microwave-oven heating. Moreover, neither microwave-oven nor autoc
lave pretreatment produced positive staining in the nuclei of Saos-2 c
ells. Therefore, tissues previously considered unsuitable for immunohi
stochemical analysis of the RB can be studied after autoclave treatmen
t.