The BlastR™ filters allows for fast and reliable preparation of genomic DNA (gDNA)-free cell lysate for western blot or immunoprecipitation when used in combination with a denaturing buffer, such as guanidine, urea or high SDS based extraction buffers that usually produce copious viscosity. gDNA contamination is a significant problem with denaturing buffers that can interfere with downstream applications like immunoprecipitation and migration of proteins in SDS-PAGE. 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.
Clarification of gDNA from BlastR™ lysate
(A) Viscous sample lysate loaded onto BlastR™ Filter.
(B) Sample passed through Filter system where gDNA is captured. >90% recovery of protein in cell 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.