Depositors Notes: 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 (email@example.com). 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.
Additional Information: For experiments involving live cells and tissue, DCAD2 supernatant is available without the antimicrobial ProClin. Contact us at 319-335-3826 or firstname.lastname@example.org for special ordering instructions.
Recommended Applications: Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blot
These hybridomas were created by your colleagues. Please acknowledge the hybridoma contributor and the Developmental Studies Hybridoma Bank (DSHB) in the Materials and Methods of your publications. Please email the citation to us.
For your Materials & Methods section:
DCAD2 was deposited to the DSHB by Uemura, T. (DSHB Hybridoma Product DCAD2)
Storage and Handling Recommendations
Although many cell products are maintained at 4°C for years without loss of activity, shelf-life at 4°C is highly variable.
To ensure retention of antibody activity, we recommend aliquotting the product into two parts:
1) a volume of antibody stored at 4°C to be used within two weeks.
2) the remaining product diluted with an equal volume of molecular grade glycerol and stored at -20°C.
While optimal Ig concentration for an application will vary, a good starting concentration for immunohistochemistry (IHC),
immunofluorescence(IF) and staining is 2-5 µg/ml.
For Western blots, the concentration is decreased by one order of magnitude (that is, 0.2-0.5 µg/ml).
All cell products contain the antimicrobial ProClin. See our FAQ section for additional information.