Selection of Detergents Suitable for IBMR3 (Mab) using Balb/c Mouse Muscle

Abstract

Monoclonal antibodies (Mab) and their fragments have been widely used for diagnostic and therapeutic purposes. Monoclonal antibodies IBMR3 hybridoma cells were produced in a previous study. In my study I used four types of detergents to fine the more suitable as the best Lysis buffer for monoclonal anti bodies using Balb/ c mouse tissue muscle. The four detergents includes; NP- 40, Igepal, Chaps and Triton X-100. Detergents were used in the laboratory to solubilize biological macromolecules such as proteins. These are none denaturing solvents; they also increase emulsification and solubilization, act as solubilize membrane proteins in their native state. The mouse samples were lysed in different lysis buffer detergents, the extracting protein where subjected on the SDS-PAGE electrophoresis, the separated protein bands were transferred to PVDF/ Polyvinylidene difluoride membrane for Immunoblotting technique. The immunblot were subsequently subjected to densitometry analysis to get the value of molecular weight, peak height and raw volume of the protein band. The results of muscle protein concentration of Blab/c mouse after using standard methods were shown (NP-40, 3.214 μg / μl), (Igepal, 3.647 μg / μl), (CHAPS, 3.925 μg / μl and Triton X-100, 4.214 μg / μl). The highest concentration of the muscle protein was obtained from using Triton X-100, followed by CHAPS, then by Igepal and in NP- 40.