Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



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Detergent Lysis of Tissue Culture Cells

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During preparation for immunoprecipitation, the lysis of cells in the presence of detergents represents a rapid method for solubilization of almost all proteins (Gilbert et al. 2002; Harlow and Lane 2006a,b; Tolosa et al. 2007; Winters et al. 2007; Ngoka 2008; Peach et al. 2012; Kan et al. 2013). Detergent-based lysis of adherent cells can be adapted to any size culture dish ranging from 96-well plates to 150-mm dishes. Suspension cultures can be collected by centrifugation or filtration for detergent-based lysis. Healthy viable cells determine the success of a cell lysis; thus, overgrown cell cultures should be avoided at all times. Cell cultures should be maintained in the logarithmic growth phase before lysis and/or addition of treatments. Upon completion of the cell lysis, cellular debris from dead cells will interfere with the immunoprecipitation by interacting with antibodies and/or beads, thus increasing nonspecific binding, and should be removed by centrifugation.

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

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