Prepare sections between 3-4micrometer thickness on Poly-L-Lysine coated slides or silanized slides to keep the samples from detaching, especially during antigen retrieval
The effect of type of incubation and two different antigen retrieval
methods on the immunodetection of Mannheimia hemolytica antigens in archived caprine lung tissues.
Second, antigen retrieval
was done by heating the sections to 95[degrees]C in 0.01 mol/L citric acid buffer (pH 6.0) for 15 min.
Slides were treated in PT-Link machine (Dako, Glostrup, Denmark), which includes deparaffinization, rehydration, and antigen retrieval
for 1 h and then cooled for 15 min.
Standardization in immunohistochemistry: the role of antigen retrieval
in molecular morphology.
These steps were followed by heating the sections in a microwave oven for antigen retrieval
using citrate buffer (pH 5.5) as previously described (Tacha and Chen, 1994).
Sections of resected specimens were cut with 3-5 um thickness and transferred to antigen retrieval
solution (Tris-EDTA Ph 9).
* Nonoptimized antigen retrieval
or use of old (weeks) tissue sections;
Under-slide heating also makes it difficult to determine the antigen retrieval
(AR) solution temperature at the tissue level, limits the volume of AR solution (which leads to evaporation issues), and makes the heaters vulnerable to corrosion and failure due to the hostile under-slide environment.
was performed by pressurized heating at 125[degrees]C in a citrate-buffered solution at pH 6.0 (Dako North America Inc., Carpinteria, CA) for Ki-67 and p63, or in an ethylenediamine tetra-acetic acid buffer solution at pH 9.0 for the estrogen receptor (ER) and progesterone receptor (PR) (Dako North America Inc., Carpinteria, CA, USA).
was performed by immersing the slides in citrate buffer (pH 6.0; 0.01 M) or EDTA (pH 8.0; 0.01 M) and subjecting them to 97[degrees]C in a water bath for 30 minutes.