Preparation of cytosolic and nuclear fractions from cultured cells and tissues is common. Cell fractionation from cultured cells is relatively easy and straightforward. However, a clear separation of cytosolic and nuclear fractions from tissues, especially frozen tissues, is challenging. The cross-contamination from frozen tissues is always an issue because of the altered tissue cellular structure caused by the freeze and thaw cycle, improper homogenization, and the use of an inadequate extraction buffer. The majority of current commercial kits are designed to treat cultured cells and tissues similarly, not paying sufficient attention to the structural difference between the two. As a result, cross-contamination of cytosolic and nuclear fractions from fresh/frozen tissues remains a major problem (see references 1 and 2 below). This novel Cytosolic and Nuclear Extraction Kit is specially designed to address the issue by employing proprietary buffers and unique protocol to minimize the cross-contamination for fresh and frozen tissue samples.
|Buffer A||25 ml|
|Buffer B||10 ml|
|Buffer D||1.5 ml|
|Buffer N||1.5 ml|
|Pestle for 1.5 ml microfuge tube||2 Units|
|Protein Extraction Powder||2 g|
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