Purification of Actin Capping Protein Antibodies Using Affinity Chromatography
Location
CSU Ballroom
Start Date
9-4-2012 10:00 AM
End Date
9-4-2012 11:30 AM
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
Antibodies are an effective tool used in many biological applications. In previous studies in the laboratory of Dr. Marilyn Hart, an antibody was generated to be used as a marker for actin capping protein in cells and tissues. Actin capping protein modulates the activity of actin monomers and filaments. The generated antibodies are a complex mixture of specific and nonspecific antibodies. Therefore, we used affinity chromatography to purify our antibody of interest from a heterogeneous group of molecules. The initial antibody mixture was passed through an affinity column to allow binding, a wash buffer was applied to remove unbound nonspecific antibodies and the elution buffer subsequently applied to release the specific antibodies. Fractions were collected and their protein concentrations determined. A fraction with an increased concentration was identified suggesting that the antibodies were purified.
Purification of Actin Capping Protein Antibodies Using Affinity Chromatography
CSU Ballroom
Antibodies are an effective tool used in many biological applications. In previous studies in the laboratory of Dr. Marilyn Hart, an antibody was generated to be used as a marker for actin capping protein in cells and tissues. Actin capping protein modulates the activity of actin monomers and filaments. The generated antibodies are a complex mixture of specific and nonspecific antibodies. Therefore, we used affinity chromatography to purify our antibody of interest from a heterogeneous group of molecules. The initial antibody mixture was passed through an affinity column to allow binding, a wash buffer was applied to remove unbound nonspecific antibodies and the elution buffer subsequently applied to release the specific antibodies. Fractions were collected and their protein concentrations determined. A fraction with an increased concentration was identified suggesting that the antibodies were purified.
Recommended Citation
Sivongxay, Sixxy. "Purification of Actin Capping Protein Antibodies Using Affinity Chromatography." Undergraduate Research Symposium, Mankato, MN, April 9, 2012.
https://cornerstone.lib.mnsu.edu/urs/2012/poster-session-A/11
