This antibody has been tested for use in ELISA, immunohistochemistry, immunofluorescence, and western blot. Specific conditions for reactivity should be optimized by the end user. Detection of Gli-3 by western blot may be enhanced if nuclear extracts are used instead of whole cell lysates as the expression/abundance of Gli-3 is likely to be low. Furthermore, Gli3 expression is likely to be developmentally regulated and induced, making it difficult to detect in whole tissue homogenates.