Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blot
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Description
Epitope: Between cadherin repeats 0 and 1. Suitable for use in immunostaining and immunoprecipitation. In western blot analysis, DCAD1, another rat monoclonal antibody to DE-cadherin, is more sensitive than DCAD2, although DCAD1 is not appropriate for staining. About requests of DCAD1, please contact Tadashi Uemura (tuemura@lif.kyoto-u.ac.jp). Conditions for immunostaining are described in our papers (Developmental Biology; Genes & Development 10, 659- 671); in addition please note the following comments. 1/ Fixatives (3.7% formaldehyde in PB or 4% paraformaldehyde in CMF & GBS containing 1mM CaCl2). Either fixation protocol works to stain embryos and discs. To stain cultured cells, only the 4% paraformaldehyde condition worked in our hands. We do not prefer any fix solutions including EDTA or EGTA, because cadherin is easily degraded under Ca++- free conditions. 2/ We suspect that the epitope may be sensitive to long exposure to alcohol. So, embryos stored in alcohol may not be stained. 3/ Photos should be taken under confocal microscope. 4/ HRP/DAB reaction has never given clear-cut signals for DE-cadherin at cell-cell contact sites. For experiments involving live cells and tissue, DCAD2 supernatant is available without the antimicrobial ProClin. Contact us at 319-335-3826 or dshb@uiowa.edu for special ordering instructions.
Immunogen
Proteins co-immunoprecipitated with an anti-alpha-catenin mAb, DCAT-1: Full length protein