Affinity Purification of an Antibody
Location
CSU North Ballroom
Start Date
25-4-2006 10:00 AM
End Date
25-4-2006 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 cytoplasmic filament that is present in all eukaryotic cells and contributes to cell shape, cell mobility, and to the organization of certain tissues such as striated muscle. Actin is regulated by accessory proteins including actin capping protein (CP) that serves to stabilize the actin filament and regulate its length. CP consists of two distinct protein subunits, an alpha (a) and a beta (p). Three a subunit isoforms (al, a2, and a3) have been identified in vertebrates. We hypothesize that the alpha subunit isoforms have distinct functions within cells and tissues. We have generated alpha isoform specific rabbit polyclonal antibodies. To purify the antibodies, I generated a DNA construct containing the gene encoding the alpha 2 protein fused to glutathione S tranferase. I overexpressed the fusion protein and am currently isolating the antibody from rabbit serum using affinity chromatography. Once the antibody is successfully isolated and purified, it can be used to investigate the location and function of the a subunit isoforms of actin capping protein in the cells and tissues.
Affinity Purification of an Antibody
CSU North Ballroom
Actin is a cytoplasmic filament that is present in all eukaryotic cells and contributes to cell shape, cell mobility, and to the organization of certain tissues such as striated muscle. Actin is regulated by accessory proteins including actin capping protein (CP) that serves to stabilize the actin filament and regulate its length. CP consists of two distinct protein subunits, an alpha (a) and a beta (p). Three a subunit isoforms (al, a2, and a3) have been identified in vertebrates. We hypothesize that the alpha subunit isoforms have distinct functions within cells and tissues. We have generated alpha isoform specific rabbit polyclonal antibodies. To purify the antibodies, I generated a DNA construct containing the gene encoding the alpha 2 protein fused to glutathione S tranferase. I overexpressed the fusion protein and am currently isolating the antibody from rabbit serum using affinity chromatography. Once the antibody is successfully isolated and purified, it can be used to investigate the location and function of the a subunit isoforms of actin capping protein in the cells and tissues.
Recommended Citation
Hart, Marilyn. "Affinity Purification of an Antibody." Undergraduate Research Symposium, Mankato, MN, April 25, 2006.
https://cornerstone.lib.mnsu.edu/urs/2006/poster-session-D/9
