Purification of Isoform Specific Actin Capping Protein Antibodies and Immunofluorescent Studies
Location
CSU 284
Start Date
27-4-2009 10:00 AM
End Date
27-4-2009 12:00 PM
Student's Major
Biological Sciences
Student's College
Science, Engineering and Technology
Mentor's Name
Marilyn Hart
Mentor's Department
Biological Sciences
Mentor's College
Science, Engineering and Technology
Description
Actin is a protein that is vital to muscle contraction and cell motility. Actin is synthesized as a monomer and polymerizes into a filament with two very distinct ends; a pointed end and a barbed end. Actin assembly is regulated by a variety of proteins including Actin Capping Protein (CP) that binds the barbed end. CP is composed of an a and a p subunit. In vertebrates, the a subunit has three isoforms: al, a2 and a3 and three beta isoforms: p 1, p2 and p3. The a and p isoforms sequences are very similar in many species, suggesting that the isoforms have specific functions. The al and a2 specific fusion proteins were prepared and expressed. The proteins were purified and used to produce isoform specific antibodies in a rabbit and a chicken. The antibodies were purified from the serum in the blood using affinity chromatography. I will evaluate the specificity of the antibodies will be evaluated using Western Blot analysis. Protein constructs were prepared, expressed, and purified the proteins. The proteins were used to generate polyclonal antibodies in chicken and rabbit. I propose to use the purified isoform specific antibodies to determine the localization of the a isoforms in murine tissues using immunofluorescence. Tissues from heart, kidney, skeletal muscle, spleen, liver, and lung, will be probed with the antibodies, and tagged with a fluorescent marker.
Purification of Isoform Specific Actin Capping Protein Antibodies and Immunofluorescent Studies
CSU 284
Actin is a protein that is vital to muscle contraction and cell motility. Actin is synthesized as a monomer and polymerizes into a filament with two very distinct ends; a pointed end and a barbed end. Actin assembly is regulated by a variety of proteins including Actin Capping Protein (CP) that binds the barbed end. CP is composed of an a and a p subunit. In vertebrates, the a subunit has three isoforms: al, a2 and a3 and three beta isoforms: p 1, p2 and p3. The a and p isoforms sequences are very similar in many species, suggesting that the isoforms have specific functions. The al and a2 specific fusion proteins were prepared and expressed. The proteins were purified and used to produce isoform specific antibodies in a rabbit and a chicken. The antibodies were purified from the serum in the blood using affinity chromatography. I will evaluate the specificity of the antibodies will be evaluated using Western Blot analysis. Protein constructs were prepared, expressed, and purified the proteins. The proteins were used to generate polyclonal antibodies in chicken and rabbit. I propose to use the purified isoform specific antibodies to determine the localization of the a isoforms in murine tissues using immunofluorescence. Tissues from heart, kidney, skeletal muscle, spleen, liver, and lung, will be probed with the antibodies, and tagged with a fluorescent marker.
Recommended Citation
Kastenschmidt, Jenna. "Purification of Isoform Specific Actin Capping Protein Antibodies and Immunofluorescent Studies." Undergraduate Research Symposium, Mankato, MN, April 27, 2009.
https://cornerstone.lib.mnsu.edu/urs/2009/oral-session-06/4
