Radio immunodiffusion

Topics: Immune system, Antibody / Pages: 4 (895 words) / Published: Nov 15th, 2013
Radial Immunodiffussion (RID)

Christian Crespo
18 October 2013
Immunology
Lab Report

Purpose of the Experiment: The objective of this experiment is to quantitatively observe the foundational reaction in our Immune system; the Antigen-Antibody interactions. The Ouchterlony procedure is what will be used in this lab to detect nature of the antibody interaction. The orientations of the band will provide more information about the interaction of antibody and antigen.
Hypothesis:
For this experiment, antibodies will be placed in wells on an agar plate, surrounded by wells containing serum (proteins, antigens etc.). If the antibody interacts with the antigen of surrounding wells, then the presence of a precipitate band will form and will further illustrate the specificity between the antibody and antigen.
Materials:
Pipette
Small vials containing antibodies or various antigens
3 Agar plates 1 sample agar plate
Straw for Making wells
Template for well making

Procedure:
To begin the Experiment, the agar plates needed to be prepared with wells. A template for the wells was provided on the lab handout and was used along with a straw to prepare the wells on the agar plate. Next, on the sample agar plates, practice runs were made by prepping sample solution in the wells to become familiar with using the transfer pipettes. After the practice run, 3 plates were prepared as follows:

The follow table demonstrates what was observed in wells AD:
Vials (AD)
Contents?
Amount?
A
Antiserum Containing Antibodies
2 Drops w/ Transfer pipette
B
Whole Serum (Protein, Antigen etc.)
2 Drops w/ Transfer pipette
C
Albumin (protein)
2 Drops w/ Transfer pipette
D
IgG
2 Drops w/ Transfer pipette

After the wells were prepared on the agar plate, they were allowed to incubate for 24-48 hours. It is important that the plates are oriented with the agar end of plate against the surface. The results



References: "Antibody-Antigen Interaction: The Ouchterlony Procedure". (n.d.). Retrieved  4, 2013, from http://www.austincc.edu/mlt/ser/OuchterlonyGelDiffuseionFall2009Edvotek.pdf Ouchterlony Double Diffusion for Antigen-Antibody Patterns - Amrita University [Video file]. (n.d.). Retrieved from http://www.youtube.com/watch?v=hmK7yYr2T54 Ouchterlony test. (n.d.). Retrieved from http://immunopaedia.org.za/index.php?id=526

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