Minute™ Total Protein Extraction Kit for Insects
Despite 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 a protein extraction point of view, the unique structure of
an exoskeleton makes it very hard to homogenize. It is also very difficult to lyse cells protectedby cuticle for
total protein extraction. Traditional solution-based protein extraction methods, such as RIPA, are inefficient
and the protein yield is low. The profile of extracted protein using traditional methods is usually incomplete.
This kit provides a highly efficient method for total protein extraction from insects using a combination of
mechanical extraction and chemical lysis. The cell lysis buffers used are much stronger than RIPA buffers.
The kit features a simple, fast single-tube protocol and optimizedbuffers for insect tissues. Researchers have
the option to choose either a denaturing or native cell lysis buffer, which is specifically tailored for insects.
The whole procedure takes less than 10 minutes to complete and the protein yield is in the range of 1-3
mg/ml. The materials provided are sufficient for 50 extractions.
Comparison of protein extraction efficiency from whole drosophila
(RIPA vs SA-07-IS)
Same amount of staring tissue and same volume of lysis buffer (200ul) were used. For RIPA buffer the tissue
was homogenized in a 1.5ml tube with a pestle for 5 minutes and incubated at RT for 10 minutes. The
homogenate was centrifuged at 10,000 x g for 10 minutes and supernatant saved. Same volume of extracted
protein was loaded in 10% SDS-PAGE and stained with Coommassie blue. For SA-07-IS denaturing
bufferwasused. Lane 1. SA-07-IS. Lane 2. RIPA buffer (R0278, Sigma)
Shipping and storage: This kit is shipped and stored at room temperature.