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.

This document is currently not available here.

Share

COinS
 
Apr 25th, 10:00 AM Apr 25th, 12:00 PM

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