A monoclonal antibody to rat liver arylsulfatase C and its application in immunohistochemistry.
We purified arylsulfatase C from rat liver microsomes and prepared a monoclonal antibody (P42C2) to the purified enzyme. By SDS-PAGE and immunoblotting analysis using P42C2, the molecular weight of the purified enzyme and of the enzyme in liver and kidney microsomes were estimated at 62,000 daltons. P42C2 caused little inhibition of arylsulfatase C activity, and was bound only slightly to liver microsomes. Localization of arylsulfatase C was studied at the light and electron microscopic level by the indirect immunoperoxidase method using P42C2. In rat liver, arylsulfatase C was detected mainly in the hepatocytes, and less frequently in endothelial cells, Kupffer's cells, and Ito's cells. In rat kidney, strong staining was observed in the straight portions of the proximal tubules. The podocytes, interstitial cells, endothelial cells, and epithelial cells of Henle's thin limbs were stained faintly. By electron microscopy, arylsulfatase C was found localized on the membranes of the endoplasmic reticulum and nuclear envelopes in these cells. These immunohistochemical findings agree with the localization demonstrated by an enzyme-histochemical method which we had previously developed.
Kawano
J
J
Department of Anatomy, Miyazaki Medical College, Japan.
Kotani
T
T
Umeki
K
K
Oinuma
T
T
Ohtaki
S
S
Aikawa
E
E
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D001192:Arylsulfatases / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D007668:Kidney / Q000032:analysis / Q000201:enzymology / Q000276:immunology; D008099:Liver / Q000032:analysis / Q000201:enzymology / Q000276:immunology*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D043266:Steryl-Sulfatase; D013429:Sulfatases / Q000276:immunology*
D000911:Antibodies, Monoclonal; D013429:Sulfatases; D001192:Arylsulfatases; D043266:Steryl-Sulfatase