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  • Polyclonal Antibody

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    Egie Arce
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    This document outlines the protocol for producing polyclonal antibodies in rabbits or mice against Streptococcus pyogenes. It involves immunizing the animals with the immunogen combined with an adjuvant, collecting pre-immune blood, and then administering additional immunizations on a schedule while collecting serum samples for testing. The protocol aims to generate an immune response through sustained antigen presentation and collecting multiple bleeds to test the antibody response over time. A final report is required.

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    This document outlines the protocol for producing polyclonal antibodies in rabbits or mice against Streptococcus pyogenes. It involves immunizing the animals with the immunogen combined with an adjuvant, collecting pre-immune blood, and then administering additional immunizations on a schedule while collecting serum samples for testing. The protocol aims to generate an immune response through sustained antigen presentation and collecting multiple bleeds to test the antibody response over time. A final report is required.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOCX, PDF, TXT or read online from Scribd
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    19 views2 pages

    Polyclonal Antibody

    Uploaded by

    Egie Arce
    This document outlines the protocol for producing polyclonal antibodies in rabbits or mice against Streptococcus pyogenes. It involves immunizing the animals with the immunogen combined with an adjuvant, collecting pre-immune blood, and then administering additional immunizations on a schedule while collecting serum samples for testing. The protocol aims to generate an immune response through sustained antigen presentation and collecting multiple bleeds to test the antibody response over time. A final report is required.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOCX, PDF, TXT or read online from Scribd
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    of 2

    Far Eastern University Finals Laboratory Experiment Animal Polyclonal Antibody Production Protocol

    1. Subject 2. Adjuvant 3. Immunogen Philippine Rabbit/Mice Complete Freunds Adjuvant/Squalene Streptococcus pyogenes. 1 mg immunogen is used for each immunization. (Peptides, proteins, bacteria, virus or any other inactivated or nontoxic organism whose antigenicity and immunogenicity have been demonstrated. Concentration advised > 1 mg / mL) The immunogen is diluted to 0.5 ml with sterile saline and combined with 0.5 ml of the appropriate adjuvant. The antigen and adjuvant are mixed thoroughly to form a stable emulsion that is injected beneath the skin of the rabbit (subcutaneously) in the area around the shoulders and intra-muscularly into the large muscle of the rear legs. (That is about 1/4 of the antigen being used in each area). This provides enhanced immune response from the sustained presentation of the immunogen. Blood is collected from the central ear artery with a 19-gauge needle and allowed to clot and retract at 37C overnight. The clotted blood is then refrigerated for 24 hours before the serum is decanted and clarified by centrifugation at 2,500 RPM for 20 minutes. Day 0 1st Immunogen Injection* Collect Preimmune Blood Sample Day 14 2nd Immunogen Injection Day 28 -3rd Immunogen Injection Collect 20 -25 mL Serum (ASO Exam) Day 35 4th Immunogen Injection Collect 20-25 mL Serum (ASO Exam) *-Pre-immune serum enables to determine whether the animals have naturally occurring antibodies that can recognize nonspecifically the antigen of interest. The rabbits can be maintained on a ten-weeks cycle of immunizations and bleeds as required by the project. The results of testing will determine whether the program should be continued, modified, or terminated. Extra bleeds can be taken upon customers requirement

    4. Procedure

    5. Bleeds

    6. Schedule

    7. Note

    8. Report

    of extended project with extra charge. This activity will be your Finals Lab Grade for Practical Exam. A paper will be required in the end of the experiment.

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