Q: SDS precipitated in the stripping buffer of pH 2.2. Will this affect the antibody stripping efficiency?

    SDS (sodium dodecyl sulfate), a common component of so-called "stripping buffer", which is applied to a membrane, to remove primary and secondary antibodies, and allow the same membrane to be used again with another antibody, is prone to precipitation at low pH, when cold. 

    Therefore, stripping buffer should be warmed up to at least 37°C before use, which will allow SDS to dissolve. Precipitated SDS will influence stripping efficiency. 

    LDS (lithium dodecyl sulfate) is soluble in the cold and at low pH, however, it will not provide the same effect as SDS for antibodies which require harsher conditions for removal from the target.

    Empirical testing is necessary in cases where most optimal stripping protocol needs to be established. 



       Antibody binding

     Note: It is important to consider, that for proteins of low abundance, the harsh stripping protocol, may also remove the target protein from the membrane, leading to increased background signal. 

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