Improved avidin—biotin—peroxidase complex (ABC) staining

Cattoretti, Giorgio; Berti, Emilio; Schirò, R; D'Amato, Lucia; Valeggio, C.; Rilke, F

doi:10.1007/bf01746607

Cited by 40 publications

(13 citation statements)

References 7 publications

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“…After washing in PBS, the sections were incubated with each primary antibody in moist boxes at 4°C overnight. Next day, the sections were rinsed in PBS at least 3 times, and further processed for immunohistochemistry by the avidin-biotin-peroxidase method [3] using a SAB-Po kit (Nichirei, Tokyo, Japan). Finally, the peroxidase activity was developed in 0.003% diaminobenzidine (DAB)/Tris-HCl (pH 7.6) solution, as described by Graham and Karnovsky [10].…”

Section: Methodsmentioning

confidence: 99%

Helicobacter pylori antigen in the glomeruli of patients with membranous nephropathy

et al. 1997

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Renal biopsy specimens from patients with membranous nephropathy (MN) were studied using immunohistochemical labelling to clarify the aetiological significance of Helicobacter pylori antigen in this disease. Sixteen specimens were examined, from 7 male and 9 female MN patients. Renal specimens from patients with diabetic nephropathy and IgA nephropathy, and from autopsied patients without renal diseases were obtained as controls. Immunohistochemical labelling was performed using one polyclonal antibody and three monoclonal antibodies against H. pylori. Specimens from 11 of the MN patients revealed granular deposits along the glomerular capillary walls, which reacted positively with polyclonal antibody after trypsin pretreatment. None of the control specimens revealed positive labelling. The MN specimens showed no positive reaction with the primary antibody, which had been treated for immunoabsorption testing using sonicated H. pylori. We also determined H. pylori status in these MN patients histologically and/or serologically. Of the 11 patients whose glomeruli were positive for anti-H. pylori antibody, 7 were suitable for analysis, and all were regarded as positive for H. pylori infection. These results suggest that the presence of a specific antigen in the glomeruli of patients with MN and H. pylori infection may be involved in the pathogenesis of MN.

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Section: Methodsmentioning

confidence: 99%

Helicobacter pylori antigen in the glomeruli of patients with membranous nephropathy

et al. 1997

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“…After two 10-min washes in PBS, a biotin-conjugated goat anti-rabbit antibody (dilution 1 : 100; Vector, Burlingame, CA) was applied for 45 min, washed twice and labeled with two rounds of modified avidin-biotin-peroxidase (ABC) complexes (Vector) [15]. The slides were then washed in PBS and developed with diaminobenzidine (Sigma, St. Louis, MO) or aminoethyl carbazole (AEC, Sigma) as previously described [15]. Endogenous peroxidase was blocked after the biotin-conjugated antibody labeling with a 30-min incubation in 0.3% hydrogen peroxide, 0.1% NaN3 in PBS.…”

Section: Immunohistochemistry and Immunofluorescencementioning

confidence: 99%

“…The slides were examined on a Zeiss (Oberkochen, FRG) epifluorescence microscope with selective FITC and TRITC filters, and photograhed on an Ilford HP5 B & W film. Ten percent formalin-fixed tissues were embedded in paraffin for morphological and immunohistochemical evaluation [15]. Mononuclear cell suspensions from BM and liver were stained for membrane antigens and nuclear TdT as previously described [9, 181. 3 Results…”

Section: Immunohistochemistry and Immunofluorescencementioning

confidence: 99%

Terminal deoxynucleotidyl transferase‐positive B cell precursors in fetal lymph nodes and extrahemopoietic tissues

et al. 1989

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Terminal deoxynucleotidyl transferase (TdT)-positive B cell precursors populate fetal lymph nodes (LN) of mid-term (16th-22th week) fetuses. These cells have a B cell phenotype (HLA-DR, CD45R, CD22, mu) and lack mature differentiation antigens (CD1c, CD20, L26). TdT+ cells are also present in the extranodal mesenteric mesenchyme, pancreas and LN primordia. Extranodal TdT+ cells are found among proliferating (i.e. Ki-67+) B cells and lack leukocyte function-associated antigen (LFA-1) and HLA-DQ; these latter molecules are acquired by nodal TdT+ elements. Adult LN, adult and fetal spleen contain less TdT+ cells than fetal LN. The fetal LN contains the full range of the B cell differentiation compartment, from the TdT+ precursor cell to the mature B lymphocyte.

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“…For immunohistochemistry [7], fixed and dried sections were incubated for 1 h at room temperature with the first antibody. Slides were washed intensively and incubated at room temperature for 30 rain with the second, biotinylated anti-mouse Ig antibody.…”

Section: Introductionmentioning

confidence: 99%

Expression of CD44 splice variants in human skin and epidermal tumours

Seiter

Tilgen

Herrmann

et al. 1996

Vichows Archiv A Pathol Anat

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Splice variants of the adhesion molecule CD44 (CD44v) are important in the lymphatic spread of rat carcinoma cells. In several human tumours expression of CD44v correlates with tumour progression. However, little is known about the physiological functions of distinct variant exons. Here we report on the immunohistological evaluation of CD44 expression in normal human skin and epidermal tumours which do not metastasise, or do so vary rarely. Frozen tissues were stained with a panel of monoclonal antibodies, recognizing epitopes of the CD44 standard isoform, as well as of variant exons v5, v6, v7, v7-v8 and v10. Stratum basale and spinosum as well as the root shaft of hairs reacted strongly with the whole panel of anti-CD44 antibodies. Stratum corneum, acinar cells of sebaceous and eccrine sweat glands stained with anti-CD44v5, anti-CD44v6 and anti-CD44v7, but not with anti-CD44v10, the latter recognizing the "epithelial isoform" (CD44v8-v10) of CD44. Ductal cells of glands and apocrine glands did not express CD44v. Compared with its expression in normal human skin, CD44v expression was reduced in basal cell carcinoma and squamous cell carcinoma of the skin. This was particularly true of CD44v10. The expression of CD44v in normal skin and dermal appendages indicates that not all combinations of variant exons are involved in tumour progression. Since the epithelial isoform is particularly downregulated in basal cell carcinoma and squamous cell carcinoma of the skin, it is unlikely that exons v8-v10 play a role in tumour progression. Rather, they may be of functional importance in maintenance of the epidermal structure.

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Improved avidin—biotin—peroxidase complex (ABC) staining

Cited by 40 publications

References 7 publications

Helicobacter pylori antigen in the glomeruli of patients with membranous nephropathy

Helicobacter pylori antigen in the glomeruli of patients with membranous nephropathy

Terminal deoxynucleotidyl transferase‐positive B cell precursors in fetal lymph nodes and extrahemopoietic tissues

Expression of CD44 splice variants in human skin and epidermal tumours

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