BlastR™ Lysis/Dilution Buffer Kit
The BlastR™ lysis buffer is a a denaturing buffer that isolates protein from all cellular compartments and produces lysates compatible with western blotting applications. When used in combination with BlastR dilution buffer it produces a cell lysate ideal for immunoprecipitation applications.
This lysis system was developed to be used with BlastR filters, which allows for fast and reliable preparation of genomic DNA (gDNA)-free cell lysate for western blot or immunoprecipitation. Unlike sonication or insulin needle gDNA shearing methods, the BlastR™ filter effectively removes gDNA contamination while having no effect on the integrity of the proteins in the lysate.
Comparison of BlastR™ versus commonly used non-denaturing and denaturing lysis buffers. Isolation of proteins from the membrane, cytoplasm, mitochondria, and nucleus.
A431 cells were lysed with BlastR™, RIPA, mPER, IP lysis, Denaturing (1% SDS), and Laemmli lysis buffers. Isolation of proteins from the membrane, cytoplasmic, mitochondrial, and nuclear markers were determined using the respective compartment marker proteins. The western blot figure highlights the ability of BlastR™ buffer to isolate proteins from all cellular compartments similarly to other denaturing buffers. Unlike ofther denaturing buffers, the BlastR™ buffer is compatible with standard protein assays, which is critical when equivalent protein loading is essential. BlastR™ buffer is also an excellent IP buffer when used with the BlastR™ dilution buffer as outlined in the datasheet. These features are outlined in the table below for easy comparison with available lysis buffers.
|D'Amico, Gabriela et al.
|ERG activity is regulated by endothelial FAK coupling with TRIM25/USP9x in vascular patterning
|Development (Cambridge, England)