Characterization of an antiserum against Atlantic cod (Gadus morhua L.) g-type lysozyme.
In this study we describe the production and characterization of an antiserum against recombinant g-type lysozyme derived from Atlantic cod. This is also the first initial analyses of g-type lysozyme protein expression in tissues of Atlantic cod. Recombinant expression and purification of cod g-type lysozyme was used for immunization to rabbit and the rabbit sera were analysed for anti g-type lysozyme antibodies using enzyme-linked immunosorbent assay (ELISA), Western blot and immunohistochemistry. ELISA results showed that antibody titres were mounted between 12,800 and 25,600 as measured at an optical density corresponding to 50% of the maximal level. By Western blot analysis the rabbit immune serum detected a single ∼23 kDa band representing the size of the injected antigen, in both spleen and head kidney homogenates from the Atlantic cod. Immunohistochemisrty detected the native folded g-type lysozyme in tissues and revealed that g-type lysozyme positive cells were observed in haematopoietic tissue of the head kidney and in red pulp of spleen. In conclusion, the rabbit anti g-type lysozyme immune sera was developed and is effectively utilized for ELISA, Western analysis as well as for immunohistochemistry. This has allowed us to obtain new knowledge about this protein regarding localization and distribution in cod tissue.
Inami
Makoto
M
Norwegian College of Fishery Science, Faculty of Biosciences, Fisheries and Economics, University of Tromsø, Tromsø, Norway. makoto.inami@uit.no
Solem
Stein Tore
ST
Jørgensen
Trond Øyvind
TØ
Larsen
Atle Noralf
AN
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D048389:Gadus morhua / Q000276:immunology*; D007106:Immune Sera / Q000096:biosynthesis* / Q000276:immunology; D007113:Immunity, Innate / Q000276:immunology; D007114:Immunization / Q000662:veterinary; D007150:Immunohistochemistry / Q000662:veterinary; D009113:Muramidase / Q000276:immunology*; D011817:Rabbits
D007106:Immune Sera; D009113:Muramidase