Wet autoclave pretreatment for antigen retrieval in diagnostic immunohistochemistry

Bànkfalvi, Àgnes; Navabi, H.; Bier, Bert; Böcker, W.; Jasani, Bharat; Schmid, Kurt Werner

doi:10.1002/path.1711740312

Cited by 189 publications

(98 citation statements)

References 7 publications

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“…Pretreatment by wet autoclaving was used for antigen retrieval. 27 The sections were incubated overnight with the polyclonal antibody against CBP. The dilution of the primary antibody was 1:100 in TBS/1% human serum.…”

Section: Immunohistochemistrymentioning

confidence: 99%

The Transcriptional Co-Activator cAMP Response Element-Binding Protein-Binding Protein Is Expressed in Prostate Cancer and Enhances Androgen- and Anti-Androgen-Induced Androgen Receptor Function

Comuzzi¹,

Lambrinidis²,

Rogatsch³

et al. 2003

The American Journal of Pathology

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Section: Immunohistochemistrymentioning

confidence: 99%

The Transcriptional Co-Activator cAMP Response Element-Binding Protein-Binding Protein Is Expressed in Prostate Cancer and Enhances Androgen- and Anti-Androgen-Induced Androgen Receptor Function

Comuzzi¹,

Lambrinidis²,

Rogatsch³

et al. 2003

The American Journal of Pathology

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“…Antigen retrieval by wet autoclave pretreatment (Bankfalvi et al, 1994), was performed, and tissue sections were subsequently immunostained with a 1:20 dilution of a monoclonal antibody against bcl-2 (mouse clone 124, kindly donated by Dr. Mason, Department of Histopathology, London University College) and with the streptavidin-peroxidase conjugate method (Shi et al, 1988). Sections incubated with mouse serum rather than specific antibody served as negative controls.…”

Section: Immunohistochemical Analysis Of Bcl-2mentioning

confidence: 99%

Analysis of SYT-SSX Fusion Transcripts and bcl-2 Expression and Phosphorylation Status in Synovial Sarcoma

Mancuso

Mezzelani

Riva

et al. 2000

Laboratory Investigation

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SUMMARY:Synovial sarcomas (SS) are characterized by a chromosomal translocation t(X;18)(p11.2;q11.2) which usually fuses the SYT gene from chromosome 18 to SSX1 or SSX2 genes on chromosome X. Also, a variant SYT-SSX4 fusion gene has recently been shown in a single SS case. In addition to these cytogenetic changes, bcl-2 expression, as assessed by immunohistochemistry, has been reported to be an almost general constitutive alteration of SS. In the present work, we analyze a series of 36 SS surgical samples (from 34 patients) by RT-PCR for the presence of the SYT-SSX1 or the SYT-SSX2 fusion transcript. The analysis was extended to SYT-SSX4 on SYT-SSX1-negative and SYT-SSX2-negative cases only. Our results showed a significant correlation between the SYT-SSX2 fusion and the monophasic SS histologic subtype. SYT-SSX1 fusion transcripts were present in both monophasic and biphasic tumors. The SYT-SSX4 fusion type was detected in a single monophasic SS. In the same series of SS cases, we also confirmed and extended the previously reported constitutive expression of bcl-2 protein, by using both immunohistochemical and western blot analysis. Moreover, we demonstrated that the BCL-2 gene is not rearranged or amplified at genomic level, indicating that the high levels of bcl-2 expression observed in SS might result from transcriptional activation of the gene and/or protein stabilization. Finally, we show that bcl-2 is not phosphorylated in tumors from patients who had been preoperatively treated with radio/chemotherapy, in tumors from untreated patients, or in an SS cell line (CME-1) after in vitro treatment with cytotoxic concentrations of DNA-damaging agents or taxanes. These data indicate that SS cells are unable to activate an apoptosis pathway involving bcl-2 phosphorylation/inactivation and may provide a possible explanation for the limited effectiveness of conventional pharmacological treatments of this tumor type. (Lab Invest 2000, 80:805-813).

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“…A 769 basepair fragment of rat p53 cDNA (Soussi et al, 1988) Immunohistochemical analysis of p53 protein Endogenous peroxidase in sections was blocked by treatment with 0.3% hydrogen peroxide in methanol for 15 min. The sections were heated at 100°C for 10 min using microwave antigen-retrieval treatment (Bankfalvi et al, 1994). Slides were immunostained using a labelled streptavidin -biotin method.…”

Section: Irradiationmentioning

confidence: 99%

Comparative alterations in p53 expression and apoptosis in the irradiated rat small and large intestine

Arai

Kida²,

Harmon³

et al. 1996

Br J Cancer

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Summary Temporal and spatial relationships between radiation-induced apoptosis and expression of p53 mRNA and protein were compared in rat small and large intestine. Apoptosis was quantified using morphological criteria, and p53 expression determined by immunohistochemistry or whole-tissue Northern analysis. In the small intestine, peak levels of apoptosis appeared earlier (4 h) than in the large intestine (6 h). p53 mRNA transcript levels in small and large intestine were not significantly altered from control levels at any time after treatment. However, in treated small and large intestine, cells showed increased positivity for p53 protein, increasing 10-fold over control levels 4-5 h after irradiation. A strong spatial relationship was found between high incidence apoptosis and p53 protein positivity. We compared published data of stem cell population positions for small and large intestine with our results. Target cells for apoptosis and p53 expression occurred at approximately fifth position from the crypt base of the small intestine, a zone coincident with stem cell population. Target cell position for apoptosis and p53 expression in the large intestine was again at fifth or sixth position from the base, but this zone is not the reported stem cell position (first or second position) for large intestine. Results from our model of radiation-induced intestinal apoptosis indicate that p53 protein is closely associated both temporally and spatially with the induction of apoptosis, and support the work of others in suggesting that p53 expression is modulated post-transcriptionally. Furthermore, our results support a hypothesis that apoptotic targeting of damaged stem cell populations, early response for apoptotic removal of DNA-damaged cells and/or early repair of these damage cells are all important parameters that determine differences in levels of tumorigenesis in the small and large intestine.

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Wet autoclave pretreatment for antigen retrieval in diagnostic immunohistochemistry

Cited by 189 publications

References 7 publications

The Transcriptional Co-Activator cAMP Response Element-Binding Protein-Binding Protein Is Expressed in Prostate Cancer and Enhances Androgen- and Anti-Androgen-Induced Androgen Receptor Function

The Transcriptional Co-Activator cAMP Response Element-Binding Protein-Binding Protein Is Expressed in Prostate Cancer and Enhances Androgen- and Anti-Androgen-Induced Androgen Receptor Function

Analysis of SYT-SSX Fusion Transcripts and bcl-2 Expression and Phosphorylation Status in Synovial Sarcoma

Comparative alterations in p53 expression and apoptosis in the irradiated rat small and large intestine

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