Depositors Notes: Flamingo #74 does not work in IP. Another mouse monoclonal antibody to Flamingo #71 is appropriate for IP experiments. About requests of #71, please contact Tadashi Uemura (email@example.com). In immunostaining pupal wings, we fix tissues with 4% paraformaldehyde (or 3.7% formaldehyde) in PBS on ice overnight (at least for 2 hours), and incubate tissues in the mAb diluted 1:10 in PBT containing 2% BSA. We do not prefer any fix solutions including EDTA or EGTA. This is because classic cadherins are degraded under Ca++- free conditions, although we have not studied how unstable Fmi is under that condition. A portion of the Fmi ectodomain (amino acid residues 539-1219) was used as immunogen.
Additional Information: Flamingo is the protocadherin-like wing polarity protein stan.
Recommended Applications: Immunofluorescence, Immunohistochemistry, 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:
Flamingo #74 was deposited to the DSHB by Uemura, T. (DSHB Hybridoma Product Flamingo #74)
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.