U-Blot™ Total Protein Extraction Kit for Insects

Despite of significant variation in body organization, insects all have the same general body structure. They have segmented bodies divided into three regions: head, thorax and abdomen. The body segments are protected by a hard exoskeleton or cuticles. From protein extraction point of view, the unique structure of exoskeleton makes it very hard to homogenize. It is also very difficult to lyse cells protected by cuticle for total protein extraction. The traditional solution-based protein extraction method such as RIPA is inefficient and protein yield is low. The profile of extracted protein using traditional method is usually incomplete. This kit provides a highly efficient method for total protein extraction from insects by a combination of mechanical extraction and chemical lysis. The cell lysis buffers used are much stronger than RIPA buffer. The kit features a simple and rapid single tube protocol and optimized buffers for insect tissues. The researchers have the option to choose either denaturing cell lysis buffer or native cell lysis buffer, which are specifically tailored for insects. The whole procedure takes less than 10 min to complete and the protein yield is in the range of 1-3 mg/ml. The materials provided are sufficient for 50 extractions.

Applications

Proteins extracted with this kit can be used for many downstream applications such as SDS-PAGE analysis, Western blotting, IP. ELISA, enzyme activity assays and proteomic analysis.The buffers are compatible with IMAC resins for his-tagged protein purification. The salts and detergents in the extracted protein sample should be removed prior to mass spectrometry analysis.