Minute™ Total Protein Extraction Kit for Tendons
A tendon/ligament is a fibrous connective tissue that attaches muscles to bone. Histologically tendons
consist of dense regular collagen fibers that are very tough and notoriously difficult to homogenize. It is also
very difficult to lyse the tendon tissues for total protein extraction. Traditional solution-based protein extraction
methods, such as RIPA, are inefficient and the protein yield is very low. Theprofile of extracted protein is also
biased (incomplete protein profile) with solution-based methods. This kit provides a highly efficient method
for total protein extraction from human or animal tendons by 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 optimized buffers for tendons. Researchers have the option to choose either a
denaturing or native cell lysis buffer, which isspecifically tailored for tendons. 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 port artery
(RIPA vs SA-04-TD)
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-04-TD denaturing
bufferwasused. Lane 1. RIPA buffer (R0278, Sigma). Lane 2. SA-04-TD.
Shipping and storage: This kit is shipped and stored at room temperature.