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Distribution of manganese superoxide dismutase in rat stomach: application of Triton X-100 and suppression of endogenous streptavidin binding activity

S Satoh, H Tatsumi, K Suzuki and N Taniguchi

Department of Anatomy, Sapporo Medical College, Japan.

The distribution of rat manganese superoxide dismutase (Mn-SOD) was immunohistochemically investigated in the rat stomach with a specific polyclonal antibody and a labeled streptavidin-biotin immunoglobulin detection system in cryosections. Parietal cells in the stomach were intensely stained, whereas the other epithelial cells in the gastric gland and pit exhibited only slight staining. Rapid-freezing and freeze- substitution immunoelectron microscopy revealed that Mn-SOD in parietal cells was mainly localized in mitochondria. Therefore, the large amount of Mn-SOD in parietal cells is due to the abundant mitochondria, in which Mn-SOD is considered to play important roles in protecting the ion pump and the cell itself from superoxide insult. Application of Triton X-100, cryosectioning, and the streptavidin-biotin system are needed to distinctly visualize Mn-SOD with our antibody. Treatment of the cryosections with Triton X-100 enhanced not only the immunoreactivity but also the false-positive staining, which showed a similar distribution pattern to that of Mn-SOD and thus made it difficult to determine the localization. The most plausible cause of the false-positive staining is thought to be endogenous biotin in the stomach, which survives paraformaldehyde fixation and is revealed by Triton X-100 treatment. Suppression of the endogenous streptavidin binding activity is important when cryosections, the streptavidin- biotin system, and Triton X-100 are employed.

Volume 40, Issue 8, pp. 1157-1163, 08/01/1992
Copyright © 1992 by The Histochemical Society


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 J. Histochem. Cytochem.Home page
M. Hollinshead, J. Sanderson, and D. J. Vaux
Anti-biotin Antibodies Offer Superior Organelle-specific Labeling of Mitochondria over Avidin or Streptavidin
J. Histochem. Cytochem., August 1, 1997; 45(8): 1053 - 1058.
[Abstract] [Full Text] [PDF]



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