pbst wash removes protein ladder western blot

The Protein A then bound detection antibodies when the immunoaffinity eluates were analyzed by Western blot. In these assays, Protein A shed from the beads, then bound to (and was eluted from) an immunoaffinity matrix designed to capture AD-related proteins. A second, unexpected artefact occurred when Protein A-conjugated Sepharose beads were used to deplete lysates of endogenous immunoglobulins prior to immunopurification of target proteins. These artefactual bands occurred in a transgene- and strain-dependent manner. First, when using biotin-avidin systems for antibody detection, endogenous biotinylated proteins created spurious bands in Western blots of brain lysates from AD patients and transgenic mouse models of AD. Here, we recount two examples of antibody-independent artefacts that have confounded the interpretation of results in our search for molecular entities associated with memory loss in Alzheimer’s disease (AD). Much attention has been paid to the potential for poorly-characterized antibodies to lead to misleading results, but antibody-independent artefacts may also occur. Antibodies are commonly used to detect or isolate proteins from biological samples.

Pbst wash removes protein ladder western blot