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.

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Apr 9th, 10:00 AM Apr 9th, 11:30 AM

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