Cells were fixed with 4% paraformaldehyde, and 0.5% Triton X-100 was used to permeabilize the cell membrane structure at room temperature. Cells were carefully removed from the cell culture plate with a fine syringe needle and placed on a slide. Slides were blocked at room temperature for 30 min by adding goat serum working solution (ZLI-9022, Zhongshan Goldenbridge Biotechnology, China) dropwise, followed by overnight incubation with diluted primary antibody, anti-Collagen II (ab34712, 1:100, Abcam) and anti-Aggrecan (13880-1-AP, 1:50, Invitrogen) at 4 °C. With addition of fluorescent secondary antibody (A32727, Invitrogen) diluted at a certain ratio, cells were further incubated for 1 h at 37 °C in a wet box. Then the nuclei with DAPI dropwise for 5 min against light. After addition of anti-fluorescence quencher dropwise, the slide was imaged under a fluorescence microscope.
Zli 9022
Manufactured by Zhongshan Golden Bridge Biotechnology
Sourced in China
The ZLI-9022 is a laboratory equipment product manufactured by Zhongshan Golden Bridge Biotechnology. It is a precision instrument designed for scientific research and analysis. The core function of the ZLI-9022 is to perform high-quality measurements and data collection in a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 5 exciter, by Zeiss (2 mentions)
Ab34712, by Abcam (1 mentions)
A32727, by Thermo Fisher Scientific (1 mentions)
Nanozoomer 2.0 rs, by Hamamatsu Photonics (1 mentions)
Pv 6002, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Zli 9018, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Ab96869, by Abcam (1 mentions)
Pv 9000, by Zhongshan Golden Bridge Biotechnology (1 mentions)
Cytopainter phalloidin ifluor 488 reagent, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Zf 0511, by Zhongshan Golden Bridge Biotechnology (1 mentions)
5 protocols using zli 9022
1
Immunofluorescent Labeling of Cartilage Markers
2
Immunohistochemical Detection of HBcAg in Liver Tissue
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Liver tissues were fixed in 4% polyformaldehyde and embedded in paraffin. Five μm tissue sections were heated at 55 °C for 2 h, deparaffinized in xylene, rehydrated in a graded series of ethanol, and then incubated in H2O2 (3%, 10 min). Tissue sections were then blocked with normal goat serum (ZLI-9022, Zhongshan Golden Bridge, Inc., Beijing, China) at 37 °C for 90 min and incubated in primary mouse anti-HBcAg (1:100, ZM-0421, Zhongshan Golden Bridge, Inc., Beijing, China) overnight at 4 °C, followed by incubation in secondary goat anti-mouse antibody (PV-6002, Zhongshan Golden Bridge, Inc., Beijing, China) for 60 min at 37 °C. Sections were then incubated with 3, 3-diaminobenzidine (DAB, ZLI-9018, Zhongshan Golden Bridge Inc., Beijing, China) for 4 min. Samples were then counterstained with hematoxylin for 4 min. Finally, sections were washed again in water, dehydrated, deparaffinized and coverslipped. For negative controls, the primary antibody was replaced by PBS to exclude false positive signals. Images were captured using the Nanozoomer slide scanner (Nanozoomer 2.0-RS; Hamamatsu Photonics, Hertfordshire, UK).
3
Immunohistochemical Analysis of Mesothelin
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Paraffin-embedded MPM specimens were cut into 5 mm sections for immunohistochemistry analysis. Sections were deparaffinized in xylene and then rehydrated with ethanol. For antigen retrieval, the specimens were incubated in a microwave oven (95 °C) in ethylene diamine tetraacetic acid (EDTA) (ZLI-9068, Zhongshan Golden Bridge Bio-technology, China) for 15 min. After natural cooling, endogenous peroxidase activity was blocked with 3% hydrogen peroxide for 15 min at room temperature. Then, sections were washed with phosphate buffer saline (PBS) for 5 min. Nonspecific antibody binding was blocked with goat serum (ZLI-9022, Zhongshan Golden Bridge Biotechnology, China) for 10 min. Subsequently, the specimens were incubated with rabbit polyclonal anti-mesothelin antibody (1:200, ab96869, abcam, UK) at 4 °C for overnight. Rewarming at 37 °C and then washed with PBS, the specimens were incubated with goat anti-rabbit secondary antibody conjugated with horseradish peroxidase (HRP) (PV-9000, Zhongshan Golden Bridge Bio-technology, China). Counter-stained with hematoxylin, the sections were rinsed, dehydrated and covered.
4
Immunofluorescence Analysis of BMSCs
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After application of tensile strain on BMSCs, BMSCs were fixed with 4% paraformaldehyde in PBS at room temperature for 15 min. After the cells were washed with PBS, the BMSCs were subsequently permeabilized with 0.1% Triton X-100 in PBS for 10 min. Then, the cells were washed with PBS again and blocked with 5% goat serum (ZLI-9022, Zhongshan Golden Bridge Biotechnology, Beijing, China) for 1 h. Then, the BMSCs were incubated with CytoPainter Phalloidin-iFluor 488 Reagent (Abcam, Cambridge, MA, USA) diluted at 1 : 1000 for 1 h at room temperature. Finally, the BMSCs were stained with DAPI (Solarbio Science & Technology Co., Ltd., Beijing, China) for 5 min at room temperature and were observed and photographed using a confocal system for imaging (LSM 5 EXCITER, Carl Zeiss, Jena, Germany).
5
Osteocalcin Immunofluorescence Staining
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Cell plating was done onto sterile glass coverslips and cultured in GM or OM for seven days. Four percent paraformaldehyde was utilized to fix the cells for 20 min at room temperature. The cells were washed (0.01 M PBS) and permeabilized (1% Triton X-100), and then they were blocked with 5% goat serum (ZLI-9022, Zhongshan Golden Bridge Biotechnology, Beijing, China) for 1 h. Primary antibody OCN (Abcam, RRID: ab13418) and anti-rabbit secondary antibody (ZF-0511, Zhongshan Golden Bridge Biotechnology, Beijing, China) were used. DAPI staining was performed to stain nuclei and then the cells were observed and photographed using a confocal system for imaging (LSM 5 EXCITER, Carl Zeiss, Jena, Germany).
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