1 / 13

Ouchterlony

Ouchterlony. Timothy G. Standish, Ph. D. Characterization of Proteins. Some proteins can be identified by their intrinsic properties - like enzyme activity - others are more difficult to characterize.

natala
Download Presentation

Ouchterlony

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Ouchterlony Timothy G. Standish, Ph. D.

  2. Characterization of Proteins • Some proteins can be identified by their intrinsic properties - like enzyme activity - others are more difficult to characterize. • Because antibodies are reasonably specific about what antigen they react or bind to, they can be used to distinguish protein antigens • The Ochterlony procedure is one of several ways in which antibodies are used to characterize proteins

  3. Antigen binding site Antigen binding site V V V V Constant Constant Constant Constant SS SS Light Chain Light Chain SS SS Antibody Structure Heavy Chains

  4. Antigen 3 Variable Heavy Variable Light Antigen Binding Antigen 1 Antigen 2

  5. IgM - A pentamer - First antibody to appear following exposure to an antigen. Because it declines rapidly in the blood, high IgM levels indicate a current infection. IgG - A monomer - Most abundant antibody in blood. IgG easily leaves the circulatory system to fight infection and crosses the placenta conferring passive immunity to a fetus. IgD - A monomer - Found on the surface of B cells probably allowing recognition of antigens thus triggering differentiation into plasma and memory B cells IgE - A monomer - The least common antibody. The tails attach to mast cells and basophils. When antigens bind, they signal release of histamine. IgA - A dimer - Produced by cells in the mucus membranes to prevent attachment of pathogens. IgA is also found in many body secretions including milk. Classes of Immunoglogulins

  6. Ouchterlony 1 2 Ab

  7. Anti Y Anti X Ouchterlony:Non-identity If wells 1 and 2 contain two separate antigens and well 3 has antibodies to both antigens, a reaction of non-identity results indicating that either X and Y are different or that well 3 has a mixture of two antibodies in it. X Y Anti X Y

  8. Non-identity:What You Really See

  9. Anti Y Anti X Ouchterlony:Partial Identity If wells 1 and 2 contain two separate antigens and well 3 has antibodies to both antigens, a reaction of partial identity may result if all antibodies react with X, but only some react with Y. indicating that X and Y are related in some way. X Y Anti X Y

  10. Non-identity:What You Really See

  11. Anti X Ouchterlony:Identity If wells 1 and 2 contain two identical antigens and well 3 has antibodies to that antigen, a reaction of identity results as all antibodies react with what is in both antigen wells indicating that the contents of both wells are closely related. X X Anti X Y

  12. Identity:What You Really See

  13. The End

More Related