RE: X-gal in paraffin

From:"Montague, Donna C"

Mary and interested others:
After fixation in aqueous aldehyde fixative of choice, rinse the tissues in deionized water followed by three rinses 10 minutes each in the following buffer:
    Prestain buffer: Final concentrations are given.
    0.1 M sodium phosphate, pH 8.3 - 8.5
    2 mM magnesium chloride
    0.1 % sodium deoxycholate
    0.2 % Triton-X 100 (or Nonidet P-40)
Stain is made up as needed in the prestain rinse solution to contain the final concentrations listed. Stain in the prepared solution 30 minutes to overnight at room temperature.
    1 mg/mL X-gal (dissolved in DMSO at 40 mg/mL, store the remaining stock at -20 oC protected from light)
    5 mM K3Fe(CN)6 (ferric cyanate)
    5 mM K4Fe(CN)6-3H2O (ferrous cyanate)
After staining, materials can be decalcified and processed as desired.
Additionally: an anti-LazZ antibody exists that works well on paraffin processed tissues following trypsin enzymatic epitope retrieval.
Hope this helps and sorry for the delay.
Donna Montague, MS
Research Associate
Center for Orthopaedic Research
UAMS Medical Center

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