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Inquiry: I will be employing your extraction buffer, which is provided in your ELISA kit, to prepare a cell lysate from a cell population.
Thereafter, i would like to quantify the total protein concentration in the cell lysate in order to normalize the concentration between samples. Does this buffer interfere with conventional protein concentration assays (Bradford or other)? In other words, will I obtain a false positive due to the buffer or the protein inhibitors present within the buffer? If so, how can I quantify my lysate after using this buffer?
Asked on Feb 02 2015
I can confirm that the extraction buffer is Triton X-100 based. It does not interfere with BCA protein method at 5X or 1X concentrations. Commercial Bradford assay specifications list Triton as interfering with the assay at 0.05%. Therefore 1X extraction buffer should be diluted at least 5-fold to be compatible with the Bradford method.
I suggest to add an extraction buffer only blank as a control measurement in your experiment.
Anja HoffmannAbcam Scientific Support
Answered on Feb 02 2015