Sensitive avidin biotin based technique for identifying liver membrane antigens in tissue sections.

Abstract

SUMMARY An enzyme immunohistochemical technique for the localisation of liver membrane antigens in tissue sections by antisera raised in guinea pigs against the liver preparation known as "liver specific membrane lipoprotein (LSP)" was developed, based on the alkaline phosphatase avidin biotin complex (ABC AP) system. Of a wide range of fixatives and fixation conditions investigated, a short (five minute) exposure of cryostat sections to Bouin's fluid provided the most satisfactory results and-together with procedures to block endogenous biotin and alkaline phosphatase-yielded clear sections with no background staining or other artefacts to interfere with specific staining patterns. The sensitivity ofthe technique approaches that ofa radioimmunoassay, as shown by the staining of the sinusoidal domains of hepatocellular plasma membranes by the guinea pig anti-LSP antisera at dilutions up to 1/50 000. Apart from its reliability and sensitivity the procedure offers additional advantages over techniques such as indirect immunofluorescence in that it provides a permanent preparation with well defined morphological details which can be seen by ordinary light microscopy.

Visualisation of capillaries in human skeletal muscle

“…Nearly all capillary endothelia in different tissues studied appeared to react strongly with UEA-I. The antibody to UEA-I and the highly sensitive ABC-AP complex (Hsu et al 1981;Sipos et al 1988), were used in the present study, provided satisfactory results. The effect of different incubation time, i.e.…”

Changes in Uterine Microvessels as a Possible Pathogenic Factor in the Development of Adenomyosis Induced by Pituitary Grafting in Mice.

“…Uterine samples embedded in paraffin were cut transversely at 4m thickness, deparafrinized, and rehydrated through graded ethanol for immunohistochemical staining using the alkaline phosphatase avidin-biotin-complex (ABC-AP) method [15,17]. To improve the staining intensity, antigen retrieval was performed by treating sections in 0.1% trypsin in 0.1% CaCl2, pH 7.8, at 37°C for 15 min.…”

Distribuição do tipo de fibras musculares e sua correlação genotípica na doença de Pompe

“…Willebrand (Sehested e Hou-Jensen 1981;Stephenson et al 1986) e seu anticorpo é altamente sensível ao complexo avidina-biotina-peroxidase (ABS) para determinar a densidade capilar (Hsu et al 1981;Sipos et al 1988) e o número de capilares em relação a diferentes tipos de fibras musculares esqueléticas humanas (Qu et al 1997).…”