1993
DOI: 10.1002/path.1711710205
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Antigen unmasking on formalin‐fixed, paraffin‐embedded tissue sections

Abstract: Enzymatic and non-enzymatic treatments for antigen unmasking on formalin-fixed, paraffin-embedded, dewaxed sections were optimized and compared by the use of a panel of antibodies of diagnostic relevance (anti-cytokeratins, vimentin, S-100, T- and B-cell receptors, Ki-67/MIB 1, muscle actin). Non-enzymatic unmasking was obtained by boiling the slides in a microwave oven in 0.01 M salt solution (pH 6) or in 6 M urea. Trypsin or pronase digestion was used for comparison and found to be necessary for some of the … Show more

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Cited by 677 publications
(347 citation statements)
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“…The IHC used to reveal the Bcl-2 protein is not always performed with the same antibody. Sometimes the protocol was performed without prior reaction of epitope unmasking on fixed issue (Cattoretti et al, 1993). To try to exclude technical biases, we performed subgroup analysis according to the most frequently used methods: IHC with antibody clone 100 and clone 124 (Figures 1 and 2).…”
Section: Discussionmentioning
confidence: 99%
“…The IHC used to reveal the Bcl-2 protein is not always performed with the same antibody. Sometimes the protocol was performed without prior reaction of epitope unmasking on fixed issue (Cattoretti et al, 1993). To try to exclude technical biases, we performed subgroup analysis according to the most frequently used methods: IHC with antibody clone 100 and clone 124 (Figures 1 and 2).…”
Section: Discussionmentioning
confidence: 99%
“…Before incubation with the primary antibodies the slides were subjected to antigen retrieval using a boiling solution of 0.01 M citrate buffer pH 6.0 (Cattoretti et al, 1993) in a microwave oven at 700 W and cooled down to room temperature in citrate buffer for 2 h. After washing in PBS, an ovemight incubation followed with mouse monoclonals anti-human ,B-catenin (1:4000, Transduction Laboratories) and anti-E-cadherin, clone HECD-1 (1:500, Zymed). Biotin-labelled rabbit anti-mouse immunoglobulins and a biotinylated HRP-streptavidin complex (both Dako) were subsequently applied for 30 min each with washes in PBS in between.…”
Section: Polymerase Chain Reaction (Pcr) Techniquementioning
confidence: 99%
“…For detection the Elite ABC kit (Vector, Burlingame, CA) was used. The immunohistochemical assay for p16 in para nembedded tissues was performed following a microwave oven antigen retrieval step using hot citrate bu er for 10 min (Cattoretti et al, 1993). Five mm para n sections of oral premalignant lesions were mounted on polysilanecoated slides, dewaxed, rehydrated and incubated with p16 antibody (normal preimmune mouse serum was used as negative control) at a 1 : 400 dilution at 48C overnight.…”
Section: Immunohistochemistrymentioning
confidence: 99%