polyclonal antibody production against mouse purified igg2a towards use in basic research

نویسندگان

zahra moradi nebrin immunology research center, faculty of medicine, tabriz university of medical sciences, tabriz, iran

jafar majidi tabriz autonomous university of medical sciences, tabriz university of medical sciences, tabriz, iran

leili aghebati maleki immunology research center, faculty of medicine, tabriz university of medical sciences, tabriz, iran

tohid kazemi department of immunology, faculty of medicine, tabriz university of medical sciences, tabriz, iran

چکیده

background: the ability of polyclonal antibodies to react with many epitopes of an antigen makes them valuable reagents in research and diagnosis. the aim of this study was purification of mouse igg2a and production of polyclonal antibody against purified mouse igg2a subclass. materials and methods: mouse igg2a was purified by proa affinity. verification method of the purified antibody was sds-page and elisa by a mouse isotyping kit. rabbit was immunized with purified igg2a. the production of antibody in rabbit was investigated by direct elisa method. rabbit serum was collected and precipitated at the final concentration of 50% ammonium sulfate. polyclonal antibody was purified by ion-exchange chromatography and labeled with hrp. the titre and cross reactivity of product was detected by direct elisa method. results: the results of sds-page in reduced and non-reduced conditions showed bands with 50-kda, 25-30 kda mw and a distinct band with 150 kda mw. isotype determination showed the presence of mouse igg2a in related fraction. the titer of anti-mouse polyclonal antibody was 200000. the optimum titer of prepared hrp conjugated igg was 4000. conjugated rabbit igg has more cross reactivity with mouse igg2b. conclusion: taking together, affinity chromatography and ion-exchange chromatography are appropriate techniques for purification of mouse igg subclasses and rabbit igg, respectively. .

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Polyclonal Antibody Production against Mouse Purified IgG2a towards Use in Basic Research

Background: The ability of polyclonal antibodies to react with many epitopes of an antigen makes them valuable reagents in research and diagnosis. The aim of this study was purification of mouse IgG2a and production of polyclonal antibody against purified mouse IgG2a subclass. Materials and Methods: Mouse IgG2a was purified by ProA affinity. Verification method of the purified antibody was S...

متن کامل

Production and Purification of a Polyclonal Antibody Against Purified Mouse IgG2b in Rabbits Towards Designing Mouse Monoclonal Isotyping Kits.

PURPOSE Mouse IgG subclasses containing IgG1, IgG2a, IgG2b and IgG3 have been defined and described both physiochemically and immunologically. METHODS Sepharose beads conjugated with protein A affinity chromatography was used for purification of mouse IgG2b. Sodium citrate buffer (0.1 M, pH: 3.5) was used for separation of mouse IgG2b. Verification of the purified fractions was monitored by S...

متن کامل

Polyclonal antibody production against bovine serum albumin conjugated artemisinin in rabbit

Abstract: The aim of the present study was to produce a polyclonal antibody against bovine serum albumin (BSA) conjugated with artemisinin. To gain an immunogenic character of artemisinin, a carboxyl group was added to it using mixed anhydride method. Then, the reactive compound of artemisinin was conjugated with BSA. The BSA+artemisinin were injected to white female New Zealand rabbits for two...

متن کامل

Production and characterization of polyclonal antibody against a synthetic peptide from β-actin protein

Objective(s):Antibodies against actin, as one of the most widely studied structural and multifunctional housekeeping proteins in eukaryotic cells, are used as internal loading controls in western blot analyses. The aim of this study was to produce polyclonal antibody against a synthetic peptide derived from N-terminal region of β-actin protein to be used as a protein loading control in western ...

متن کامل

Production and purification of polyclonal antibody against F(ab')2 fragment of human immunoglobulin G

Antibodies are essential tools of biomedical and biochemical researches. Polyclonal antibodies are produced against different epitopes of antigens. Purified F(ab')2 can be used for animal’s immunization to produce polyclonal antibodies. Human immunoglobulin G (IgG) was purified by ion exchange chromatography method. In all stages verification method of the purified antibodies was sod...

متن کامل

A methodological approach for production and purification of polyclonal ‎antibody against dog IgG ‎

Antibodies are a class of biomolecules that has an important role in the immune system and lots of applications in biotechnological methods and in pharmaceutics. Production and purification of antibodies in laboratory animals is one of the first ways to manufacture of these prominent tools. The obtained antibodies from these process could be used in various types of bioassay techniques such as ...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
research in molecular medicine

جلد ۴، شماره ۱، صفحات ۳۰-۳۵

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023