RE: X-gal in paraffin
Message
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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