Cat # WA-014
Manual & Protocol | Material Safety Data Sheets (MSDS)
The traditional method for nuclei isolation from animal cells/tissues depends on the use of non-ionic detergent to lyse the plasma membrane and release nuclei which can be subsequently isolated by low-speed centrifugation and further purified by density gradient. One major problem associated with the traditional method is the clumping of nuclei after isolation, which, in some applications such as DNA and RNA extraction, may not be a problem. However, in other applications such as single nucleus sequencing or proteomics, clumping of isolated nuclei represents a major problem. The anti-clumping nuclei storage buffer is designed to address this issue. Nuclei resuspended in this buffer show significantly reduced clumping as compared to those resuspended in PBS with 5% BSA. The nucleus suspension can be stored at 4oC for days without significant aggregation and change in morphology. The nuclei can also be frozen in the buffer at -80oC or in dry ice for transportation or long-term storage.
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Comparison of the Storage Buffer with 5% BSA in PBS for Anti-Clumping Efficacy. Nuclei of frozen mouse spleen and lung tissues were isolated by Nucleus Isolation Kit for Tissues/cells (Cat# SN-047, Invent Biotechnologies, Plymouth MN). The isolated nuclei pellets were resuspended in 0.4 ml nuclei storage buffer or 0.4 ml 5% BSA in PBS and incubated at 4oC overnight. The nuclei were resuspended by vortexing briefly and stained with trypan blue to compare the nuclear morphologies.
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