The primary-culture astrocytes were grown on glass coverslips. After stimulation with or without the inflammatory media for 48 h, the cells were harvested and fixed in 4% paraformaldehyde and blocked in blocking buffer at room temperature for 1 h, followed by incubation in the primary anti-rat IL-6 (PeproTech) or rabbit anti-mouse IL-6 (Bioss Ltd., Beijing, China) or anti-GFAP antibodies at 4 °C overnight. The cells were then incubated with the FITC-conjugated goat anti-mouse secondary antibody (BD, Franklin Lakes, NJ, USA) or goat anti-rabbit secondary antibodies (red, Beyotime Ltd., Shanghai, China) at room temperature for 1 h in the dark. The cells were then incubated with 2 μg/mL of 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI; Roche) for 10 min before the slides were mounted. The images were obtained and analyzed with a fluorescence microscope (Nikon 80i) with a cold CCD camera (Nikon DS-Ri1) and NIS-Elements F 3.0 software. For cryostat sections, two independent observers evaluated 30 optical fields for every one of 10 sections per animal (7 animals per group) with a qualitative scale.
2 4 amidinophenyl 6 indolecarbamidine dihydrochloride dapi
Manufactured by Roche
Sourced in Germany, United States
2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) is a fluorescent stain used for detecting and quantifying nucleic acids, particularly DNA. It binds to the minor groove of double-stranded DNA, emitting blue fluorescence upon excitation. DAPI is commonly used in various biological and biochemical applications, such as cell and tissue staining, fluorescence microscopy, and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Ds ri1, by National Instruments (1 mentions)
Fitc conjugated goat anti mouse secondary antibody, by BD (1 mentions)
Chondroitin sulfate a sodium salt cs, by Merck Group (1 mentions)
Rat anti mouse cd31 antibody, by BD (1 mentions)
Iba1 antibody, by Fujifilm (1 mentions)
Fluorescein isothiocyanate isomer 1 fitc, by Thermo Fisher Scientific (1 mentions)
5 protocols using 2 4 amidinophenyl 6 indolecarbamidine dihydrochloride dapi
1
Immunofluorescence Analysis of Astrocyte IL-6 Expression
2
Cell Morphology on Nano-Scaffolds
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F-actin staining was used to define the cell morphology on scaffolds with different nanotopographies, which were well-oriented or random nanofibres. Aligned/random dishes (diameter 10 mm) were placed into a 48-well plate, and PDLSCs were seeded at a density of 2 × 104 cells per well. On the 3rd and 7th day, cell-scaffold complexes were rinsed with PBS once, and then fixed with 4% paraformaldehyde for 10 min and permeated with 0.5% Triton X-100 for 5 min. Next, 100 nM rhodamine phallolidin (Cytoskeleton, USA) was added to each well and worked in a dark environment for 30 min, after which 100 nM 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) (Roche, Germany) was added, working for 30 seconds. Finally, all staining reagents were removed completely and after enough rinsing the samples were observed by LCSM. For SEM observation of cells planted on the surfaces of AL and RD, samples at day 3 and 7 were obtained and subjected to general processing.
By determining the long axis of the nucleus and actin fibres of each cell, the angulations of cell orientation were calculated with ImageJ. Three random views of each group in high magnification were selected.
By determining the long axis of the nucleus and actin fibres of each cell, the angulations of cell orientation were calculated with ImageJ. Three random views of each group in high magnification were selected.
3
Chondroitin Sulfate-Based Biomaterial Synthesis
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Chondroitin sulfate A sodium salt (CS) (Type A 70%, balanced with Type C, from bovine trachea), glycidyl methacrylate (GMA), poly(ethylene glycol) methyl ether (MPEG, Mn = 2,000), ε-caprolactone (ε-CL), acryloyl chloride (AC), triethylamine (TEA) and ammonium persulfate (98%) (APS) were purchased from Sigma-Aldrich Company, USA. 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) was obtained from Roche, USA. The rest of the reagents were used as received. All the reagents were analytic grade quality.
4
Characterization of Diesel Particulate Matter
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Standard reference material (SRM) 2975 diesel particulate matter (industrial forklift) was obtained from the National Institute for Standards and Technology (Gaithersburg, MD, USA). 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) was purchased from Roche Diagnostics (Indianapolis, IN, USA). The rabbit anti-ionized calcium-binding adaptor molecule 1 (IBA-1) antibody was purchased from Wako (Richmond, VA, USA), the rat anti-mouse CD31 antibody from BD Biosciences (Franklin Lakes, NJ, USA), and the mouse anti-rat CD31 from Invitrogen (Carlsbad, CA, USA). LPS (strain O111:B4) was purchased from EMD Chemicals (Gibbstown, NJ, USA). All other reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA).
5
Synthesis and Characterization of Polymeric Nanomaterials
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mPEG (Mn = 500, 1900, and 5000) was purchased
from Alfa Aesar (U.K.), and PCL diols (Mn = 2000) was purchased from
Dow Chemical (USA). Both of them were dehydrated under vacuum before
use. Paclitaxel and Taxol were obtained from Jinhe Biotechnology Co.,
Ltd. (Shanghai, China) and West China Hospital, Sichuan University,
respectively. Fluorescein isothiocyanate isomer I (FITC, 90%) was
purchased from Acros Organics (USA). 2-(4-Amidinophenyl)-6-indolecarbamidine
dihydrochloride (DAPI) was obtained from Roche Diagnostics (Germany).l -Cystine diethyl ester was purchased from Shanghai Bajiu Industry
Co., Ltd. (China), and H-Lys(Boc)-OH and Phe-OMe were purchased from
Sichuan Anpaibo Biotechnology Co., Ltd. (China). Dicyclohexylcarbodiimide
(DCC) and N-hydroxysuccinimide (HOSU) were purchased
from Shanghai Aladdin Biochemical Technology Co., Ltd. (China). CCK-8
was purchased from Tongren Institute of Chemistry (Japan).
from Alfa Aesar (U.K.), and PCL diols (Mn = 2000) was purchased from
Dow Chemical (USA). Both of them were dehydrated under vacuum before
use. Paclitaxel and Taxol were obtained from Jinhe Biotechnology Co.,
Ltd. (Shanghai, China) and West China Hospital, Sichuan University,
respectively. Fluorescein isothiocyanate isomer I (FITC, 90%) was
purchased from Acros Organics (USA). 2-(4-Amidinophenyl)-6-indolecarbamidine
dihydrochloride (DAPI) was obtained from Roche Diagnostics (Germany).
Co., Ltd. (China), and H-Lys(Boc)-OH and Phe-OMe were purchased from
Sichuan Anpaibo Biotechnology Co., Ltd. (China). Dicyclohexylcarbodiimide
(DCC) and N-hydroxysuccinimide (HOSU) were purchased
from Shanghai Aladdin Biochemical Technology Co., Ltd. (China). CCK-8
was purchased from Tongren Institute of Chemistry (Japan).
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