جستجوی مقالات مرتبط با کلیدواژه « Western-Blot » در نشریات گروه « دامپزشکی »

Partially fed (4 -5 days) H. a. anatolicum (Iranian isolate) female adult ticks were used for preparation of antigens of midgut which are, the gut supernate antigen (GS), gut membrane antigen (GM), triton extracted gut membrane antigen (TX-GM) and sodium dodecyl sulphate treated Triton extracted gut membrane antigen (SDS-TX-GM). The New Zealand white rabbits were immunized with GS, GM, TX- GM and SDS-TX-GM antigens. The first inoculation on day 0 was administered after emulsification with Freund`s complete adjuvant and second inoculation was given on day 14 with incomplete Freund`s adjuvant and third inoculation was given on day 28 with incomplete Freund`s adjuvant. Blood drawn was from the marginal ear vein on day 38. The humoral immune response was studied by Enzyme Linked Immunosorbent Assay (ELISA) and antibody titers in the sera of immunized rabbits on 10 days after the last booster dose of antigen was 1/32000. After electrophoresis of purified midgut antigens (GS, GM, TX-GM, SDS-TX-GM) 4µg of each were applied on a 10% acryl amide gel and blotted to nitrocellulose paper for reaction with its concern immune sera. By analysis of this Western blot, we got bands of 110, 95, 85, 66, 59, 49, 42 Kda for GSAg, 95, 85, 66, 49, 42 Kda for GM Ag, 66, 49 Kda for TX-GMAg and 66Kda for SDS-TX-GMAg responsible for induction of resistance in the host. The prepared antigens showed engorged nymphs ranged from 32-41%, 29-56%, 29-49% and 29-47% in response to GS, GM, TX-GM and SDS-TX-GM antigens respectively. Immunodominant antigenic proteins of 66 Kda were detected in the sera of rabbits immunized by all above antigens which can be candidate for single immunogen if in future studies show acceptable results. The immunized rabbits were challenged, 10 days after the last booster dose of antigens, with 700 larvae of H. a. anatolicum ticks by releasing them on one ear of rabbits and maximum protection in rabbits against H. a. anatolicum was induced with the one immunized with GSAg.

To detect the tumor antigens in BLV associated lymph nodes and comparing with FLK-BLV cell culture antigens by immunoblotting. Design: Observational study. Samples: Nineteen BLV lnfected and 2 healthy lymph nodes. Procedure: SDS-PAGE and Westem blotting were car:ried out for all infected and non infected lymph nodes.

SDS-PAGE results revealed no considerable differences between infected and non-infected lymph nodes. Among all protein profiles of BLV infected lymph nodes, in western blot test, protein bands with 54,52,47, 46, 44, 43,39 and 36 kD weight were detected to be immunogenic.BothT2 and 57 kD bands were exist in all infected and non-infected lymph nodes as well as FLKBLV cells. Cells infected by BLV (FLK-BLV) demonstrated only 48 kD band as an immunoreactive protein identical to only one sample profile.

The present work demonstrates at least 8 different immunogenic proteins in BLV infected lymph nodes that could be classified to three distinct profiies. 48kD protein demonstrated in FLK- BLV and just one of the profiles, seems to be a viral antigen that is expressing in certain conditions.