Anti–Lucifer Yellow Dye, Anti–Alexa Fluor 405/Cascade Blue Dye and Anti–Alexa Fluor 488 Dye Antibodies—Note 14.1

Molecular Probes anti–lucifer yellow dye antibodies were specifically developed to overcome certain limitations of lucifer yellow CH (L453, L682, L12926, L1177). Lucifer yellow CH is an aldehyde-fixable fluorescent cell tracer that has long been used by neuroscientists to identify patterns of gap junctional communication, to assay the outgrowth of developing neurons and to characterize the morphology of neurons from which electrical recordings have been made. Even though the cell soma of a lucifer yellow CH–filled neuron may be brightly stained, its finer processes can sometimes be faint and may fade rapidly or be obscured by the more intensely stained portions of the neuron. Investigators have been able to overcome these limitations by using anti–lucifer yellow dye antibodies in conjunction with standard enzyme-mediated immunohistochemical methods to develop a more permanent, fade-free signal for light microscopy. Anti–lucifer yellow dye antibodies have also been used to develop tissue for electron microscopy and to distinguish neurons filled with lucifer yellow CH from those injected with the lectin Phaseolus vulgaris leucoagglutinin (PHA-L). Molecular Probes Cascade Blue hydrazide (C3239), which can also be fixed in place with aldehyde-based fixatives, can potentially be used as a second label with lucifer yellow CH to characterize the morphology of interacting neurons.

For these applications,we offer unconjugated and biotinylated rabbit polyclonal anti–lucifer yellow dye (A5750, A5751) and anti–Alexa Fluor 405/Cascade Blue dye (A5760) antibodies. Similarly, our rabbit polyclonal anti–Alexa Fluor 488 dye antibody (A11094) can be used to detect Alexa Fluor 488 hydrazide (A10436), Alexa Fluor 488 hydroxylamine (A30629), Alexa Fluor 488 cadaverine (A30676) and Alexa Fluor 488 biocytin (A12924) in fixed-cell preparations. See Anti-Dye and Anti-Hapten Antibodies—Section 7.4 (Anti-fluorophore and anti-hapten antibodies—Table 7.8) for a complete description of these and other anti-fluorophore and anti-biotin antibodies.