Brugia Malayi - Clark Science Center - Smith College

HSP25 Immunoprecipitation from C2C12 Skeletal Muscle Cells
Claire Maesner
Immunoprecipitation is a technique that utilizes the high affinity of an antibody for an antigen to purify a target protein from a
highly complex solution, even a cell culture extract. This strategy was used to specifically isolate heat shock protein 25, a molecular
chaperone protein involved in the cellular stress protection mechanisms, out of cells from the C2C12 murine skeletal muscle cell
line. A direct immunoprecipitation was accomplished employing an anti-HSP25 antibody to isolate the desired protein. Running
the sample on a one dimensional SDS polyacrylamide gel confirmed the isolation of the target had been successful (Figure 1,
lanes 1-3 within the red box). A purified HSP25 standard is in lane 5, after the molecular weight ladder in lane 4. Following this
accomplishment, the immunoprecipitation technique is being modified to pull down and identify the target’s associated proteins,
thus further revealing its physical interactions with other proteins within these skeletal muscle cells. (Supported by the Howard
Hughes Medical Institute)
Advisor: Stylianos P Scordilis
2012
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