Monoclonal antibodies that inhibit the enzyme activity of NAD+-dependent 15-hydroxyprostaglandin dehydrogenase

Abstract

[13] administration. These studies suggest that an understanding of the regulation of this enzyme activity and of its intracellular abundance could shed new light on the function of the enzyme and of prostaglandins. Such studies would be greatly aided by the production of antibodies against the enzyme. Questions about the distribution, abundance and turnover of the enzyme under various conditions could be examined by measuring the quantities of the enzyme protein and with immunocytochemical techniques. The extent of structural analogy of the enzyme from different sources could also be probed by site-specific monoclonal antibodies. In this report, we describe our success in obtaining three monoclonal antibodies to the enzyme, each of which inhibits the enzyme to a different degree, suggesting that these monoclonal antibodies all interact with the enzyme near the active site, but in different

Induction of NAD+-Linked 15-Hydroxyprostaglandin Dehydrogenase Expression by Androgens in Human Prostate Cancer Cells

“…PGE 2 was supplied by Cayman Chemical Co. ECL ϩ plus Western Blotting Detection System RPN 2132 was obtained from Amersham Pharmacia Biotech. Rabbit antiserum against human placental 15-PGDH was generated as described previously (7). Rabbit antisera against human COX-1 (LLPPLPVLLADPGAPTPV) and COX-2 (NASSSRSGLDDINPTVLLK) specific sequences were generated using glutathione-S-transferase fusion protein as antigens (8).…”

15-Hydroxyprostaglandin Dehydrogenase Is Down-regulated in Gastric Cancer

“…Specimens were immunostained with a rabbit antiserum against human 15-PGDH (1:5,000; ref. 24) or a COX-2 -specific anti-human mouse monoclonal antibody (1:200 Cayman).…”

“…As primary antibodies rabbit antiserum against human 15-PGDH (1:10,000; ref. 24), goat polyclonal anti -human-COX-2 (1:1,000), or anti -h-actin (1:500, both Santa Cruz) were used. Secondary antibodies were goat anti-rabbit (1:2,000; Dako) or donkey anti-goat (1:2,000; Santa Cruz).…”

Localization of 15-Hydroxy Prostaglandin Dehydrogenase in Human Fetal Membranes, Decidua, and Placenta during Pregnancy

“…To determine the specificity of IR-PGDH staining, sections were incubated in antibody dilution buffer in the absence of primary anti body, after substitution of the primary' antibody with non-immunc rabbit serum, or with primary antibody that had been preabsorbed with purified excess type I PGDH for 18 h at 4 °C [ 11 ]. As a positive control we included a section of term fetal membranes in every batch of slides that was processed.…”

“…The sections were stained for IR-PGDH using the avidin-biotin procedure (Vector ABC; Vector Lab oratories, Burlingame, Calif.,) as described previously [9], The poly clonal primary antibody (Cayman Laboratories, Ann Arbor, Mich.) was raised in rabbits against purified human placental type I PGDH [11] and used at a dilution of 1:2,000 and 1:3.000. Additional sec tions were stained for cytokeratin as a marker of epithelial and trophoblast cells, using a polyclonal primary antibody (anti-keratin, wide spectrum screening; 1:1,500 and 1:2,000, Dako Corporation, Santa Barbara, Calif.) raised in rabbits against bovine muzzle epider mal keratin.…”