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Should I Order an Affinity Purified Antibody?
The use of an affinity purified antibody, versus using raw serum or an IgG fraction, is dependent upon several factors. First, the use of an affinity purified antibody insures that the antibody in question is targeting only the sequence in question. Even though 21st Century Biochemicals uses SPF (specific pathogen-free) rabbits, animals produce antibodies to various proteins throughout their life. Thus, as would be expected, affinity purified antibodies tend to have lower backgrounds, lower nonspecific binding, and thus give cleaner signals. This is very important in specific immunological applications such as immunostaining or immunocytochemistry. The use of affinity purified antibodies is strongly recommended in such cases. Further, the production of phosphospecific antibodies requires that the serum undergo two rounds of purification. The first is the purification of antibodies that recognize the phosphopeptide, followed by a depletion step whereby the antibodies that bind to the peptide without regard to the presence of the target phosphoamino acid are removed. Thus phosphospecific antibodies are always affinity purified.
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