The line blot assay: Problems with titrating first and second antibodies for Western blot and immunohistochemistry assays?

Authors: Rojas‐Espinosa, O.; Silva‐Miranda, M.; Wek‐Rodriguez, K.; Arce‐Paredes, P.

Source: Scandinavian Journal of Clinical and Laboratory Investigation, Volume 66, Number 4, June 2006 , pp. 357-362(6)

Publisher: Informa Healthcare

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Abstract:

We describe a technique designed to assess the optimal dilution of primary and secondary antibodies, to be used in Western blot, dot blot, the multi‐antigen print immunoassay (MAPIA) and immunohistochemistry assays. The method that we call “line blot” is not an alternative but a practical, complementary tool for the above techniques that assures definitive results are obtained from single assays, so there is no need to repeat the assay. As with most immunoenzymatic assays, the line blot assay is very sensitive, allowing the detection of absolute amounts of antigen as low as 2.5 ng in the 0.5 cm‐long segment line (see Results), depending on the strength of the secondary, enzyme‐labelled antibody.

Keywords: Dot blot; ELISA; immunohistochemistry; MAPIA; Western blot

Document Type: Research article

DOI: http://dx.doi.org/10.1080/00365510600672809

Affiliations: 1: Department of Immunology, National School of Biological Sciences, National Polytechnic Institute, México D.F., México

Publication date: 2006-06-01

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