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Expression of constitutive heat shock protein-70 in normal (non-stressed) rabbit urinary bladder tissue
Authors:Yang Zhao  Robert M. Levin  Frederick C. Monson  Samuel Chacko
Affiliation:(1) Division of Urology, University of Pennsylvania, 3006 Courtyard Building 3400 Spruce St, 19104 Philadelphia, Pennsylvania, USA;(2) Department of Pathobiology, University of Pennsylvania, 3006 Courtyard Building 3400 Spruce St, 19104 Philadelphia, Pennsylvania, USA
Abstract:The expression of constitutive HSP-70 in the urinary bladder was determined by SDS-PAGE and western blotting using a mouse monoclonal antibody against HSP-70. The western blot analysis showed that the mouse anti-HSP-70 cross-reacted with a 70 kDa protein present in the extracts of the urinary bladder muscle and mucosa. Densitometric scanning of the western blots allowed us to specifically quantitate the relative amounts of the HSP-70. The quantitation of the HSP-70 by combining immunoblotting and densitometry using a laser scanner is reproducible and this technique requires only a small amount of tissue. The amounts of HSP-70 can be estimated from a standard curve of nanogram(ng) of HSP-70 vs absorption from the immunoblots. The amounts of HSP-70 in the muscular and mucosal layers in the body of the urinary bladder are more than those in the base of the bladder. The presence of HSP-70 in the muscle and mucosal epithelium of the bladder was demonstrated by immunohistochemical analysis of freshly removed tissue from the base and the body of bladder from normal animals.
Keywords:heat shock protein 70 (HSP-70)  urinary bladder smooth muscle  immunoblotting  immunofluorescence  laser densitometry
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