Event Title

Purification of Isoform Specific Actin Capping Protein Antibodies and Immunofluorescent Studies

Presenter Information

Jenna Kastenschmidt, Minnesota State University, Mankato

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.