The cells were fixed with 4% paraformaldehyde (AS1018, Aspen, Wuhan) for 20 min. Then, the fixed cells were blocked with 10% bovine serum albumin (AS1018, Aspen, Wuhan) for 30 min, followed by incubation at 4°C overnight with the following primary antibodies: anti-E-cadherin (20874-1-AP, San Ying, Wuhan), anti-N-cadherin (Abcam, Ab18203), and anti-β-catenin (20874-1-AP, San Ying, Wuhan). After incubating the cells with CY3- or FITC-conjugated goat anti–mouse or goat anti–rabbit IgG (AS-1111, Aspen, Wuhan) secondary antibody at 37°C for 40 min, they were captured using fluorescence microscopy.
As1018
Manufactured by Aspen
Sourced in China
The AS1018 is a laboratory equipment designed for general use. It features a compact and durable construction, allowing for reliable performance in various laboratory settings. The core function of the AS1018 is to provide a versatile platform for a range of laboratory tasks.
Automatically generated - may contain errors
4 protocols using as1018
1
Immunofluorescence Staining of Cell-Cell Junctions
2
Immunofluorescence Staining of Cell Markers
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We fixed the cells by incubating in 4% PFA solution (AS1018, Aspen, Wuhan) for 30 min. Then, after blocking the fixed cells with 10% BSA (10735078001, Roche) at 37 °C for 30 min, they were incubated at 4 °C overnight with the following primary antibodies: anti-SERPINH1 (ab109117, Abcam); anti-E-cadherin (20874-1-AP, San Ying, Wuhan); and anti-N-cadherin (Abcam, Ab18203). Then, the cells were incubated with CY3- or FITC-conjugated goat anti–mouse or goat anti–rabbit IgG (AS-1111, Aspen, Wuhan) at 37 °C for 1 h. The cells were imaged and analyzed under a fluorescence microscope.
3
Histochemical Staining of Cardiac Fibrosis
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The heart tissues were washed with saline solution (10092-18; Jinuo Co, Hangzhou, China), fixed in 4% paraformaldehyde (As1018; Aspen Biological; ≥24 h at 37°C), and then processed using analytical grade ethanol and xylene. The paraffinized sections (4–5 µm thick) were stained with H&E (hematoxylin, 3–8 min at 37°C and eosin, 1–3 min at 37°C) to visualize inflammatory cell infiltration, cardiomyocyte necrosis and apoptosis, and fibrosis in myocardial tissue. As an efficacious histochemical stain for collagen detection in tissue sections, Picrosirius red (PSR; AS1067; Aspen Biological was used to differentiate the collagen types. In addition, using Masson's trichrome staining (1–3 min at 37°C), the collagen volume fraction (CVF), and the interstitial collagen volume fraction were examined to quantify the degree of fibrosis. Cardiomyocytes were stained red and fibrous tissues were stained in blue by Masson's Trichrome staining. The sections were then examined by light microscopy and images were captured at ×40 and ×200 magnification. The percentages of fibrosis in the heart were analyzed using Image-Pro Plus software (version 6.0; Media Cybernetics, Inc., Rockville, MD, USA) and the captured images and their average values were assessed.
4
Evaluating Apoptosis in Human ES A673 Cells
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Human ES A673 cells were seeded onto sterile coverslips (10212432C; Citotest Labware Manufacturing Co., Ltd, Jiangsu, China), which were inserted in 6-well plates (5 × 105 cells/well). 48 h after the transfection with 15 nM nCAR/miR-34a-5p, control RNA or vehicle, the coverslips were fixed with 4% paraformaldehyde (AS1018; Aspen, Wuhan, China). After being permeabilized and blocked, the coverslips were incubated with primary antibody against cleaved-caspase-3 (1:200; AF7022; Affbiotech, OH, USA) at 4°C overnight, and then incubated with FITC labeled goat anti-rabbit secondary antibody (1:50; AS-1110; Aspen, Wuhan, China). Nucleus was counterstained with DAPI (AS1075; Aspen, Wuhan, China). Images were taken and observed with a fluorescence microscope (IX51; Olympus Corporation, Shinjuku, Tokyo, Japan). Cleaved-caspase-3-positive cells were counted under the fluorescence microscope for percentages of positive stained cells which were compared between different groups of treatments (38 (link), 39 (link)).
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