Microwave-assisted antigen retrieval and incubation with cox-2 antibody of archival paraffin-embedded human oligodendroglioma and astrocytomas

Temel, Şehime Gülsün; Minbay, Fatma; Kahveci, Zeynep; Jennes, Lothar

doi:10.1016/j.jneumeth.2006.02.025

Cited by 11 publications

(6 citation statements)

References 37 publications

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“…Antigen retrieval is needed to achieve the optimal immunostaining for cox-2, since the staining without AR resulted in poor immunoreaction (Temel et al 2006). In order to get a better and specific staining, antigen retrieval method was employed: Sections were mounted on poly-Llysine-coated slides.…”

Section: Antigen Retrievalmentioning

confidence: 99%

Cyclooxygenase-2 expression in astrocytes and microglia in human oligodendroglioma and astrocytoma

Temel

Kahveci

2009

J Mol Hist

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Cyclooxygenases (cox) are potent mediators of inflammation and two cox-isoenzymes, cox-1, cox-2, are described to date. Cox-2 is cytokine-inducible in inflammatory cells and enhanced cox-2 expression has been attributed a key role in the development of edema and immunomodulation in pathologically altered brain tissues. In normal cerebral cortex cox-2 is present only in neurons, but not in the glial or vascular endothelial cells. The function of microglia in glioma biology is unclear. Microglia have both neurotrophic and neurotoxic functions and have been shown to release a variety of cytokines. Our preliminary results showed that the expression pattern of cox-2 is predominantly neuronal although glial expression was observed with the correlation of high malignancy. In this study we aimed to assess the phenotypes (astrocyte, microglia) of the cox-2-expressing glial cells in various types of human gliomas and to compare their expression patterns. For this purpose we employed dual immunohistochemistry for cox-2 and GFAP (astrocyte) or LCA-MAC (microglia-macrophage) in archival formalin-fixed, paraffin embedded human tissue diagnosed as oligodendroglioma and/or astrocytoma. The results showed that cox-2 immunoreactivity is up-regulated in the neurons according to the tumor grade. Most of the cox-2 immunoreactive glia were GFAP-positive in anaplastic oligodendrogliomas and at lesser extend in glioblastomas. Cox-2 and LCA co-localization was detected in more glial cells in glioblastomas. It may be speculated that the induction of cox-2 in microglia may contribute to the deleterious effects of prostanoids in cerebral edema formation during the progression of oligodendrogliomas. The detection of cox-2 in astrocytes surrounding the necrotic areas might be important to develop new strategies, such as the usage of cox-2 inhibitors combine with chemotherapy and radiotherapy in the treatment of glioma patients.

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Section: Antigen Retrievalmentioning

confidence: 99%

Cyclooxygenase-2 expression in astrocytes and microglia in human oligodendroglioma and astrocytoma

Temel

Kahveci

2009

J Mol Hist

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“…It has, therefore, become common practice to increase antigen exposure (a protocol known as antigen retrieval) by using microwave irradiation, so that antibodies have enhanced binding capabilities to target antigens. Several in-depth studies into antigen retrieval and fixation of tissues using microwave incubation have been reported (Patterson & Bulard, 1980;Hafajee & Leong, 2004;Emersen et al, 2006;Temel et al, 2006). These investigations state that either by a pulsed microwave exposure prior to or during incubation of the sample with the primary antibody, staining times can be drastically reduced from many hours to minutes.…”

Section: B Immunohistochemical Staining and Tissue Fixationmentioning

confidence: 99%

“…Just a few years later, microwave technology began to emerge in protocols of a more bio-analytical nature. Such preliminary techniques included the exposure of antigens for immunohistochemical staining in embedded tissue samples (Patterson & Bulard, 1980;Hafajee & Leong, 2004;Emersen et al, 2006;Temel et al, 2006), increased acid hydrolysis of proteins for amino acid analysis (Gilman & Woodward, 1990;Chiou & Wang, 1990;Chen, Chiou, & Wang, 1991;Davidson, 1996), increased catalysis of enzymatic and chemical cleavages for peptide mapping (Zhong, Marcus, & Li, 2005;Hua, Low, & Sze, 2005), and increased removal of PTMs for improved protein characterization (Lee et al, 2005a,b;Sandoval et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Microwave‐assisted proteomics

Lill¹,

Ingle²,

Liu³

et al. 2007

Mass Spectrometry Reviews

145

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State-of-the-art proteomic analysis has recently undergone a rapid evolution; with more high-throughput analytical instrumentation and informatic tools available, sample preparation is becoming one of the rate-limiting steps in protein characterization workflows. Recently several protocols have appeared in the literature that employ microwave irradiation as a tool for the preparation of biological samples for subsequent mass spectrometric characterization. Techniques for microwave-assisted biocatalyzed reactions (including sample reduction and alkylation, enzymatic and chemical digestion, removal and analysis of posttranslational modifications and characterization of enzymes and protein-interaction sites) are described. This review summarizes the various approaches undertaken, instrumentation employed, and reduction in overall experimental time observed when microwave assistance is applied. #

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“…Epitope retrieval was achieved by heating the sections in DAKO target retrieval solution (high pH) for 12 minutes. 18 Endogenous peroxidase was blocked with 3% H 2 O 2 for half an hour. After 2 washes in phosphate-buffered saline, slides were incubated with DAKO protein block serum-free for 1 hour to block unspecific staining.…”

Section: Rna Extraction and Real-time Reverse Transcriptionypolymerasmentioning

confidence: 99%

Alternative Splicing-Related Factor YT521

Zhang

Hausen

Orlowska-Volk

et al. 2010

International Journal of Gynecological Cancer

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Microwave-assisted antigen retrieval and incubation with cox-2 antibody of archival paraffin-embedded human oligodendroglioma and astrocytomas

Cited by 11 publications

References 37 publications

Cyclooxygenase-2 expression in astrocytes and microglia in human oligodendroglioma and astrocytoma

Cyclooxygenase-2 expression in astrocytes and microglia in human oligodendroglioma and astrocytoma

Microwave‐assisted proteomics

Alternative Splicing-Related Factor YT521

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