Antibody Purifications and Antibody Labeling
Once an adequate polyclonal response has been reached, the crude serum can be purified by two distinct methods:
- Protein A (or G) affinity chromatography is used for purifying all the IgGs present in the sample. For better results and to enrich for specific IgG antisera, we recommend the elimination of antibodies with low affinity and specificity but higher binding capacity, such as IgM by purification of the antiserum.
- Immunoaffinity purification is recommended for purifying antigen specific antibodies from a pool of polyclonal antibodies, principally when they are to be used in immunocytochemistry or in immunofluorescence, where a high background would interfere with analysis. In this procedure, the pure antigen (free peptide) is covalently bound to a solid support: CNBr-Sepharose.The specific antibodies within the pool bind the antigen-sepharose, while the aspecific antibodies flow through the column. Finally the specific antibodies are eluted from the column and collected in TRIS buffer; their positivity and titer are reconfirmed by ELISA testing.
Purified antibodies can be labeled by Primm with fluorochromes or biotin. There are different ways of labeling antibodies. Fluorescent labels are essential for some techniques, such as immunocytochemistry or cell sorting. Biotinylated antibodies can be detected using labeled streptavidin or avidin, which have a high affinity for biotin and show low levels of background binding.
At Primm we label antibodies produced by our staff, as well as antibodies supplied by the customer.
Custom polyclonal antibodies
Phospho specific antibodies
Phospho specific antibody
Custom polyclonal antibody
Custom polyclonal Abs