Strategies for improving the immunohistochemical staining of various intranuclear prognostic markers in formalin-paraffin sections: Androgen receptor, estrogen receptor, progesterone receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen revealed by antigen retrieval techniques

Taylor, Clive R.; Shi, Shan; Chaiwun, Benjaporn; Young, Lillian; Imam, S. Ashraf; Côté, Richard J.

doi:10.1016/0046-8177(94)90198-8

Cited by 249 publications

(104 citation statements)

References 20 publications

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“…Briefly, formalin fixed, paraffin-embedded sections of 5µm thickness were deparaffinized with xylene and subsequently rehydrated in descending gradient of ethanol and water. Antigenic unmasking was performed by microwaving the sections in antigen retrieval solution (100 mM Glycine-HCl, and 10 mM EDTA, pH 3.0) as described previously [12][13][14]. Endogenous peroxidase activity was blocked by incubating sections for 10 min with 3% hydrogen peroxide and further non-specific background was blocked by incubating with 10% goat serum and 0.1% Triton X100 in PBS for 2 h at room temperature.…”

Section: Immunohistological Analysis Of Pxr In Liver Tissue Sectionsmentioning

confidence: 99%

“…8B). Antigenic epitope unmasking appears to be one of the critical factors that govern the development of signal in immunohistochemistry [12][13][14]. Hence, we used a series of target antigen retrieval buffers (Glycine-HCl buffer pH 3.0, Citrate buffer pH 6.0 and Tris buffer pH 9.0, along with 10 mM EDTA) during micro-waving the tissue sections.…”

Section: Anti-pxrmentioning

confidence: 99%

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Purification of full-length human Pregnane and Xenobiotic Receptor: polyclonal antibody preparation for immunological characterization

et al. 2005

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Pregnane and Xenobiotic Receptor (PXR; or Steroid and Xenobiotic Receptor, SXR), a new member of the nuclear receptor superfamily, is thought to modulate a network of genes that are involved in xenobiotic metabolism and elimination. To further explore the role of PXR in body's homeostatic mechanisms, we for the first time, report successful prokaryotic expression and purification of full-length PXR and preparation of polyclonal antibody against the whole protein. The full-length cDNA encoding a 434 amino acids protein was sub-cloned into prokaryotic expression vector, pET-30b and transformed into E. coli BL21(DE3) cells for efficient over expression. The inclusion body fraction, containing the expressed recombinant protein, was purified first by solubilizing in sarcosine extraction buffer and then by affinity column chromatography using Ni-NTA His-Bind matrix. The efficacy of anti-PXR antibody was confirmed by immunocytology, Western blot analysis, EMSA and immunohistochemistry. The antibody obtained was capable of detecting human and mouse PXR with high specificity and sensitivity. Immunofluorescence staining of COS-1 cells transfected with human or mouse PXR showed a clear nuclear localization. Results from immunohistochemistry showed that level of PXR in liver sections is immunologically detectable in the nuclei. Similar to exogenously transfected PXR, Western blot analysis of cell extract from HepG2 and COLO320DM cells revealed a major protein band for endogenous PXR having the expected molecular weight of 50 kDa. Relevance of other immunodetectable bands with reference to PXR isoforms and current testimony are evaluated. Advantages of antibody raised against full-length PXR protein for functional characterization of receptor is discussed and its application for clinical purposes is envisaged.

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Section: Immunohistological Analysis Of Pxr In Liver Tissue Sectionsmentioning

confidence: 99%

Section: Anti-pxrmentioning

confidence: 99%

Purification of full-length human Pregnane and Xenobiotic Receptor: polyclonal antibody preparation for immunological characterization

et al. 2005

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“…Five-m serial sections were treated with xylene to remove paraffin, rehydrated, treated with 3% hydrogen peroxide in methanol to eliminate endogenous peroxidase activity, and treated for 15 minutes in a microwave oven 32 with 0.05 mol/L glycine-HCl, pH 3.5, 33 for antigen retrieval. The sections were then incubated with 5% normal horse serum with 0.01% Triton X in phosphate-buffered saline (PBS) at pH 7.4 for 20 minutes to eliminate nonspecific background immunostaining.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Expression of Lysosome-Associated Membrane Proteins in Human Colorectal Neoplasms and Inflammatory Diseases

Furuta

Ikeda

Nakayama

et al. 2001

The American Journal of Pathology

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The lysosome-associated membrane proteins (LAMPs)-1 and -2 are major constituents of the lysosomal membrane. These molecules are known to be among the most glycosylated proteins of several types of cells and cancer cells, and their expression in cancer cells is marked by a distinct difference in the structures of the oligosaccharides as compared to nonmalignant cells. We analyzed by immunohistochemistry the intensity and distribution of LAMP-1 and LAMP-2 in 9 human colorectal cancer cases and in 16 control cases, including inflammatory diseases (diverticulitis, ulcerative colitis, and Crohn's disease). LAMP proteins were expressed more intensely in the epithelium of colorectal neoplasms than in normal mucosa (P < 0.05), and no significant differences were found between adenoma and cancer cells (P > 0.05) in the same tissue section.

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“…False negatives of hormone receptor are not uncommon, especially with increasing time delay before tissue fixation, increased duration of fixation, and type of fixative, all of which may have been the case in our patient cohort. False negatives increase especially in BC with a low receptor positivity such as is typically seen in patients of African descent [23][24][25][26]. At present, it is impossible to say whether this played a part in our study population, but the lack of IHC to determine predictive markers is bound to hamper treatment decision making, and possibly outcome.…”

Section: Survival In Monthsmentioning

confidence: 99%

A nationwide analysis of incidence and outcome of breast cancer in the country of Surinam, during 1994–2003

Leeuwaarde

Vrede

Henar

et al. 2011

Breast Cancer Res Treat

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In this study, we describe the incidence, treatment, and outcome of breast cancer (BC) during the period 1994-2003 in the South-American country of Surinam and compare these with those of BC in the Netherlands. Pathology reports and hospital charts from all BC cases diagnosed between 1994 and 2004 were retrieved from Surinam's single pathology laboratory and its five hospitals. Data on demographics, tumor characteristics, treatment, and follow-up were gathered. We compared our data to BC statistics of first generation immigrants from Surinam to the Netherlands. 421 patients were diagnosed with BC during the study period. The age-adjusted incidence rate was 26 per 100,000 compared to 65/100,000 in first generation Surinamese women in the Netherlands. The majority had a fairly advanced stage at presentation, with 60% of tumors larger than 2 cm, and 41.6% with lymph node involvement. Because of the absence of radiotherapy facilities, local treatment in most patients was radical mastectomy. Adjuvant hormonal therapy (51.6%) was administered more frequently than adjuvant chemotherapy (20.3%). A significant number of patients were lost to follow-up, resulting in a median follow-up duration of only 23 months. The 5-year overall survival was 79%. BC incidence in Surinam is low compared to that in the western world, but the advanced stage at diagnosis, the low utilization of systemic adjuvant therapy, and the inadequate follow-up may lead to poor outcomes. A number of steps are underway to improve the level of cancer care in Surinam.

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Strategies for improving the immunohistochemical staining of various intranuclear prognostic markers in formalin-paraffin sections: Androgen receptor, estrogen receptor, progesterone receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen revealed by antigen retrieval techniques

Cited by 249 publications

References 20 publications

Purification of full-length human Pregnane and Xenobiotic Receptor: polyclonal antibody preparation for immunological characterization

Purification of full-length human Pregnane and Xenobiotic Receptor: polyclonal antibody preparation for immunological characterization

Expression of Lysosome-Associated Membrane Proteins in Human Colorectal Neoplasms and Inflammatory Diseases

A nationwide analysis of incidence and outcome of breast cancer in the country of Surinam, during 1994–2003

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