Immunohistochemistry:Article Title: Nuclear Klf4 accumulation is associated with cetuximab drug-resistance and predicts poor prognosis of nasopharyngeal carcinoma
Article Snippet: .. Immunohistochemistry (IHC) method After deparaffinization and rehydration, the paraffin-embedded sections were subjected to antigen retrieval for 2 min. Then, the slides were incubated overnight at 4 °C with the following primary antibodies: (dilution 1:200, abcam, MA). .. Serial tumor sections and IHC staining evaluated the expression of proteins.
Article Title: An activin receptor IIA ligand trap corrects ineffective erythropoiesis in β-thalassemia
Article Snippet: Immunohistology.Paraffin-embedded bone marrow or spleen sections (5 μm) were stained with H & E for morphological analysis. .. For immunohistochemistry and confocal analysis, sections were processed for antigen retrieval as indicated by each manufacturer and incubated overnight with primary antibodies against GDF11 proform (ab71347, Abcam; 1/100 dilution), mature GDF11 (sc28910, Santa Cruz; 1/20 dilution), activin A (AF338, R & D Systems; 1/100 dilution), activin B (MAB659, R & D Systems; 1/100 dilution), activin receptor II (sc-25451, Santa Cruz; 1/100 dilution), phospho-Smad1/5 (#9511, Cell Signaling; 1/100 dilution), phospho-Smad2 (#3101, Cell Signaling; 1/100 dilution), Ter-119 (#553673, BD Biosciences; 1/100 dilution), CD71 (136800, Life Technologies; 1/200 dilution) or F4/80 (sc-377009, Santa Cruz Biotechnology; 1/50 dilution). .. Following incubation with biotinylated species-specific secondary antibodies (1/250 dilution) for 1 h and streptavidin-HRP for 30 min (Vectastain, ABC kit; Vector Laboratories, Burlingame, USA), the immunoperoxidase reaction was visualized by addition of 3,3′-diaminobenzidine (DAB) in chromagen solution (Dako, Carpinteria, USA).
Article Title: Ang-2 promotes lung cancer metastasis by increasing epithelial-mesenchymal transition
Article Snippet: For analysis of cell migration, we use the modified Boyden Chambers without the Transwell-precoated Matrigel membrane filter with the method performed as above. .. Immunohistochemistry (IHC) TMA slides used for IHC analysis were deparaffinized, and eroxidase was quenched with methanol and 3% H2 O2 for 15 min. For antigen retrieval, the sections were boiled under pressure in citrate buffer (pH 6.0) for 3 min and then incubated for 120min with primary mouse anti-human Ang-2 (ab155106, Abcam, USA), washing with phosphate buffered saline (PBS), incubated with horse reddish peroxidase (HRP)-conjugated goat anti-mouse IgG (A21020; Abbkine, USA) for 15 min at 1:5000 dilution, and washed again with PBS. .. In finally, the slides were incubated with diaminobenzidine and counterstained with hematoxylin solution, dehydrated in ethanol, cleared in xylene, and cover-slipped.
Incubation:Article Title: Nuclear Klf4 accumulation is associated with cetuximab drug-resistance and predicts poor prognosis of nasopharyngeal carcinoma
Article Snippet: .. Immunohistochemistry (IHC) method After deparaffinization and rehydration, the paraffin-embedded sections were subjected to antigen retrieval for 2 min. Then, the slides were incubated overnight at 4 °C with the following primary antibodies: (dilution 1:200, abcam, MA). .. Serial tumor sections and IHC staining evaluated the expression of proteins.
Article Title: An activin receptor IIA ligand trap corrects ineffective erythropoiesis in β-thalassemia
Article Snippet: Immunohistology.Paraffin-embedded bone marrow or spleen sections (5 μm) were stained with H & E for morphological analysis. .. For immunohistochemistry and confocal analysis, sections were processed for antigen retrieval as indicated by each manufacturer and incubated overnight with primary antibodies against GDF11 proform (ab71347, Abcam; 1/100 dilution), mature GDF11 (sc28910, Santa Cruz; 1/20 dilution), activin A (AF338, R & D Systems; 1/100 dilution), activin B (MAB659, R & D Systems; 1/100 dilution), activin receptor II (sc-25451, Santa Cruz; 1/100 dilution), phospho-Smad1/5 (#9511, Cell Signaling; 1/100 dilution), phospho-Smad2 (#3101, Cell Signaling; 1/100 dilution), Ter-119 (#553673, BD Biosciences; 1/100 dilution), CD71 (136800, Life Technologies; 1/200 dilution) or F4/80 (sc-377009, Santa Cruz Biotechnology; 1/50 dilution). .. Following incubation with biotinylated species-specific secondary antibodies (1/250 dilution) for 1 h and streptavidin-HRP for 30 min (Vectastain, ABC kit; Vector Laboratories, Burlingame, USA), the immunoperoxidase reaction was visualized by addition of 3,3′-diaminobenzidine (DAB) in chromagen solution (Dako, Carpinteria, USA).
Article Title: Ang-2 promotes lung cancer metastasis by increasing epithelial-mesenchymal transition
Article Snippet: For analysis of cell migration, we use the modified Boyden Chambers without the Transwell-precoated Matrigel membrane filter with the method performed as above. .. Immunohistochemistry (IHC) TMA slides used for IHC analysis were deparaffinized, and eroxidase was quenched with methanol and 3% H2 O2 for 15 min. For antigen retrieval, the sections were boiled under pressure in citrate buffer (pH 6.0) for 3 min and then incubated for 120min with primary mouse anti-human Ang-2 (ab155106, Abcam, USA), washing with phosphate buffered saline (PBS), incubated with horse reddish peroxidase (HRP)-conjugated goat anti-mouse IgG (A21020; Abbkine, USA) for 15 min at 1:5000 dilution, and washed again with PBS. .. In finally, the slides were incubated with diaminobenzidine and counterstained with hematoxylin solution, dehydrated in ethanol, cleared in xylene, and cover-slipped.
Article Title: Expression and prognostic significance of MYL9 in esophageal squamous cell carcinoma
Article Snippet: Immunohistochemistry (IHC)Formalin-fixed, paraffin-embedded samples were cut into 5-μm sections and processed for IHC. .. Tissue sections prepared for antigen retrieval by microwave treatment in EDTA buffer (pH 9.0) were incubated with primary antibodies against MYL9 (1:1000; ab191393, Abcam) and α-smooth muscle actin (SMA) (1:500; Zhongshan Bio-Tech, Zhongshan, China). .. Immunostaining was performed using the EnVision System with diaminobenzidine (Dako Cytomation, Glostrup, Denmark).
Article Title: Hereditary pancreatitis model by blastocyst complementation in mouse
Article Snippet: The paraffin-embedded sections were de-paraffinized in Hemo-De (Farma, Japan) and were rehydrated in a graded series of ethanol. .. The slides were heated in an antigen retrieval buffer for 40 minutes, blocked with goat or horse serum for 20 minutes at room temperature, and incubated with monoclonal mouse anti-Gfp antigen antibody (1:1000, Abcam, England) or monoclonal mouse anti-PTF1A antigen antibody (1:50, BD Biosciences, US) overnight at 4°C. .. The Vectastain ABC System (Vectastain, Funakoshi, Japan) was used to visualize the antigens.
Article Title: Leptin System in Obese Dog Skin: A Pilot Study
Article Snippet: Immunohistochemistry Immunohistochemistry was carried out as follows [ ]: sections were rehydrated and dipped for 10 min in 3% H2 O2 to reduce endogenous peroxidase activity. .. To perform antigen retrieval, sections were microwaved for 15 min in 10 mM citric acid (pH 6.0) or incubated with Historeveal (Abcam, Cambridge, UK) for 5 min as anti-LEPR antibody regards. .. The sections were then blocked with 1:10 normal serum ( ) for 30 min and incubated at room temperature with the primary antibody ( ) for 1h (for anti-CD20, Anti-CD3 and anti-Iba-1 antibodies) or overnight (for anti-LEP and anti-LEPR antibodies).
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