Proper use of Stripping Buffer for Western Blotting
Western blotting is widely used by the scientific community. Most western blotting is performed manually with relatively inexpensive equipment. The quality of WB in publications and subsequent analysis and interpretation of the data can be variable, which can lead to inconsistent result. This may be due to poor laboratory technique or the lack of knowledge of the critical steps involved in WB. Proper quality control procedures should be in place to produce reproducible results and robust data generation. The selection of reagents for WB is also an important step, let’s take for example, stripping buffer (cat#SB200). The proper use of stripping buffer for WB can be overlooked. Precision Biosystems stripping buffer has been optimized for efficient and fast removal of primary and secondary antibodies leaving the antigen attached to the membrane and ready for the next immunodetection. Precision stripping buffer can be used during manual or automated blot processing and works for both PVDF and nitrocellulose membranes. Based on our studies, Precision stripping buffer works well with for both low and high affinity antibodies. During automated blot processing with the BlotCycler Touch or Mini, stripping can be combined with new antibody incubation. Membranes can be stripped and re-probed up to 8 times without significant loss of sample protein. Our stripping buffer also allows for the elimination of the blocking step; this in conjunction with an automated blot processing instrument (BlotCycler) eliminates user error, provides reproducible results, increases sensitivity, and publication quality results time after time.
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