Fluorescence In Situ Hybridization:Article Title: Notch2 Controls Prolactin and Insulin-Like Growth Factor Binding Protein-1 Expression in Decidualizing Human Stromal Cells of Early Pregnancy
Article Snippet: Immunofluorescence First trimester decidual tissue samples (n = 5) were fixed with 7.5% formaldehyde and embedded in paraffin (Merck, Darmstadt, Germany). .. Deparaffinized tissue sections (3 µm) were boiled in 1x Target Retrieval Solution, pH 6.1 (Dako, Glostrup, Denmark) and blocked with 0.05% fish skin gelatin (Sigma Aldrich), followed by incubation overnight at 4°C with the following primary antibodies: Notch1 (1∶100; Cell Signaling Technology, 3608), Notch2 (1∶100; Cell Signaling Technology, 5732), Notch3 (2 µg/ml, Santa Cruz Biotechnology, sc-5593), Notch4 (5 µg/ml; Lifespan, LS-C137135), Jagged1 (2 µg/ml; Santa Cruz Biotechnology, sc-8303), Jagged2 (1∶100; Cell Signaling Technology, 2210), DLL1 (10 µg/ml; Abcam, ab76655), DLL3 (2 µg/ml; Santa Cruz Biotechnology, sc-67269), DLL4 (17.6 µg/ml; Abcam, ab-7280), vimentin (1.44 µg/ml; GeneTex, 100619 or 1∶200, DAKO, M7020), CD45 (1∶100, DAKO, M0701), cytokeratin 7 (1.96 µg/ml; DAKO, M7018) and HLA-G1 (1∶200, Exbio, 1P-292-C100). .. Subsequently, sections were incubated with goat anti-mouse or anti-rabbit IgG conjugated to Alexa Fluor 488 or Alexa Fluor 568 (2 µg/ml, Molecular Probes, Life Technologies) for 1 hour at room temperature, counterstained with DAPI (1 mg/ml, Roche Diagnostics, Mannheim, Germany) and mounted in Fluoromount-G (SouthernBiotech, Birmingham, AL).
Incubation:Article Title: Notch2 Controls Prolactin and Insulin-Like Growth Factor Binding Protein-1 Expression in Decidualizing Human Stromal Cells of Early Pregnancy
Article Snippet: Immunofluorescence First trimester decidual tissue samples (n = 5) were fixed with 7.5% formaldehyde and embedded in paraffin (Merck, Darmstadt, Germany). .. Deparaffinized tissue sections (3 µm) were boiled in 1x Target Retrieval Solution, pH 6.1 (Dako, Glostrup, Denmark) and blocked with 0.05% fish skin gelatin (Sigma Aldrich), followed by incubation overnight at 4°C with the following primary antibodies: Notch1 (1∶100; Cell Signaling Technology, 3608), Notch2 (1∶100; Cell Signaling Technology, 5732), Notch3 (2 µg/ml, Santa Cruz Biotechnology, sc-5593), Notch4 (5 µg/ml; Lifespan, LS-C137135), Jagged1 (2 µg/ml; Santa Cruz Biotechnology, sc-8303), Jagged2 (1∶100; Cell Signaling Technology, 2210), DLL1 (10 µg/ml; Abcam, ab76655), DLL3 (2 µg/ml; Santa Cruz Biotechnology, sc-67269), DLL4 (17.6 µg/ml; Abcam, ab-7280), vimentin (1.44 µg/ml; GeneTex, 100619 or 1∶200, DAKO, M7020), CD45 (1∶100, DAKO, M0701), cytokeratin 7 (1.96 µg/ml; DAKO, M7018) and HLA-G1 (1∶200, Exbio, 1P-292-C100). .. Subsequently, sections were incubated with goat anti-mouse or anti-rabbit IgG conjugated to Alexa Fluor 488 or Alexa Fluor 568 (2 µg/ml, Molecular Probes, Life Technologies) for 1 hour at room temperature, counterstained with DAPI (1 mg/ml, Roche Diagnostics, Mannheim, Germany) and mounted in Fluoromount-G (SouthernBiotech, Birmingham, AL).
Article Title: Primary Thymic Extranodal Marginal-Zone B-Cell Lymphoma of Mucosa-Associated Lymphoid Tissue Type Exhibits Distinctive Clinicopathological and Molecular Features
Article Snippet: The immunohistochemical study was performed using a streptavidin-biotin method as described elsewhere. .. Deparaffinized tissue sections were treated with 3% hydrogen peroxide for 5 minutes, washed in phosphate-buffered saline, and then incubated with antibodies to the following antigens: cytokeratin (AE1/AE3; DAKO, Kyoto, Japan), CD20 (L26, DAKO), CD3 (polyclonal, DAKO), CD23 (1B12; Novocastra, Newcastle-on-Tyne, England), CD43 (MT1; Bio-Science, Emmerbrucke, Switzerland), CD5 (4C7, Novocastra), immunoglobulin (Ig) A (polyclonal, DAKO), IgD (polyclonal, DAKO), IgG (polyclonal, DAKO), IgM (polyclonal, DAKO), κ and λ light chains (polyclonal, DAKO), bcl2 protein (124, DAKO), cyclin D1 protein (5D4; MBL, Nagoya Japan), and ALK protein (polyclonal; Nichirei, Tokyo, Japan). .. Slides were incubated with biotinylated secondary antibody and then with peroxidase-streptavidin conjugate, followed by reaction with diaminobenzidine-hydrogen peroxide.
Generated:Article Title: Thrombospondin-1 is a critical effector of oncosuppressive activity of sst2 somatostatin receptor on pancreatic cancer
Article Snippet: Tumor xenografts were started with the s.c. inoculation of BxPC-3/mock or/sst2 cells into athymic female mice. .. Deparaffinized tissue sections of TMA or of tumor xenografts were placed in citrate buffer, pH 6.0, heated in microwave oven for 3 × 5 min, probed with TSP-1 (NeoMarkers) or sst2 antibody (generated in our laboratory), and then with horseradish peroxidase-conjugated secondary antibody (Dako). .. A solution of 3-amino-9-ethylcarbazole (AEC) was used as chromogen (Dako), and sections were counterstained with hematoxylin.
In Situ Hybridization:Article Title: Etiological factors in primary hepatic B-cell lymphoma
Article Snippet: Antibody, clone, and source were as follows: CD3 (PS1, Novocastra, UK), CD5 (4C7, Novocastra), CD10 (56C6, Novocastra), CD20 (L26, Dako, USA), CD23 (1B23, Novocastra), IL2α receptor (CD25, 4C9, Novocastra), CD30 (BerH2, Dako), Bcl1 (5D4, IBL, Japan), Bcl2 (124, Dako), Bcl6 (P1F6, Novocastra), multiple myeloma 1 (MUM1p, Dako), anti-HBs antigen (ZCH16, Nichirei, Japan), and IgG4 (HP6026, Zymed, USA). .. In situ hybridization for detection of Epstein–Barr virus-encoded RNA (EBER) Deparaffinized tissue sections were digested with proteinase K and hybridized in a solution of 50% formamide containing fluorescein isothiocyanate (FITC)-labeled EBER oligonucleotides using a Dako hybridization kit (DakoCytomation, Glostrup, Denmark). .. Hybridization products were detected using rabbit anti-FITC antibody, then incubated with ChemMate Envision and colored with DAB.
Hybridization:Article Title: Etiological factors in primary hepatic B-cell lymphoma
Article Snippet: Antibody, clone, and source were as follows: CD3 (PS1, Novocastra, UK), CD5 (4C7, Novocastra), CD10 (56C6, Novocastra), CD20 (L26, Dako, USA), CD23 (1B23, Novocastra), IL2α receptor (CD25, 4C9, Novocastra), CD30 (BerH2, Dako), Bcl1 (5D4, IBL, Japan), Bcl2 (124, Dako), Bcl6 (P1F6, Novocastra), multiple myeloma 1 (MUM1p, Dako), anti-HBs antigen (ZCH16, Nichirei, Japan), and IgG4 (HP6026, Zymed, USA). .. In situ hybridization for detection of Epstein–Barr virus-encoded RNA (EBER) Deparaffinized tissue sections were digested with proteinase K and hybridized in a solution of 50% formamide containing fluorescein isothiocyanate (FITC)-labeled EBER oligonucleotides using a Dako hybridization kit (DakoCytomation, Glostrup, Denmark). .. Hybridization products were detected using rabbit anti-FITC antibody, then incubated with ChemMate Envision and colored with DAB.
Immunohistochemistry:Article Title: Incidental Collision Tumor of Hepatocellular Carcinoma and Neuroendocrine Carcinoma
Article Snippet: Methods The liver specimen was fixed in 10% buffered formalin, embedded in paraffin, cut into 4-μm-thick sections, and stained with hematoxylin and eosin. .. Immunohistochemistry was performed on deparaffinized tissue sections using the avidin-biotin-peroxidase method and a Dako AutoStainer (Carpinteria, CA, USA). .. After antigen retrieval, the following panel of primary antibodies was applied: Hep Par1 (hepatocellular antigen, 1:50; Dako), CD34 (1:200; Dako), synaptophysin (1:10; Dako), chromogranin (1:50; Dako), CK7 (1:1,500; Dako), CK19 (1:50; Dako), CD56 (1:50; Vision BioSystem), MIB-1 (1:100; Dako) and β-Catenin (1:250; Dako).
Avidin-Biotin Assay:Article Title: Incidental Collision Tumor of Hepatocellular Carcinoma and Neuroendocrine Carcinoma
Article Snippet: Methods The liver specimen was fixed in 10% buffered formalin, embedded in paraffin, cut into 4-μm-thick sections, and stained with hematoxylin and eosin. .. Immunohistochemistry was performed on deparaffinized tissue sections using the avidin-biotin-peroxidase method and a Dako AutoStainer (Carpinteria, CA, USA). .. After antigen retrieval, the following panel of primary antibodies was applied: Hep Par1 (hepatocellular antigen, 1:50; Dako), CD34 (1:200; Dako), synaptophysin (1:10; Dako), chromogranin (1:50; Dako), CK7 (1:1,500; Dako), CK19 (1:50; Dako), CD56 (1:50; Vision BioSystem), MIB-1 (1:100; Dako) and β-Catenin (1:250; Dako).
Staining:Article Title: Exercise delays allogeneic tumor growth and reduces intratumoral inflammation and vascularization
Article Snippet: Any cell that was of the size 6–9 μm in diameter with a round, densely stained nucleus and a relatively small amount of pale basophilic, nongranular cytoplasm was counted as a lymphocyte. .. Deparaffinized tissue sections of 3-μm thickness were stained with antibody against von Willebrand factor VIII-associated antigen (Dako, Carpinteria, CA) and counterstained with hematoxylin. .. Factor VIII-associated antigen-stained slides were analyzed on days 5 and 10 to verify the results of the visual determination of blood-vessel density in tumors from six mice per group at each time point.
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