Recombinant porcine IFN-α was obtained as a gift sample from Harbin Veterinary Institute of Chinese Academy of Sciences. Caco-2 cell line (Human colon carcinoma) was purchased from Stem Cell Bank, Chinese Academy of Sciences. High Glucose DMEM and Hank's balanced salt solution (HBSS) were obtained from Gibco, USA. HOOKTM-Dye Labeling Kit was purchased from Biosciences, USA. ALP Kit was purchased from Jiancheng Bioengineering institute, Nanjing, China. Trypsin-EDTA, penicillin and streptomycin solution, glutamine and non-essential amino acids all were obtained from Hyclone, USA. Fetal bovine serum (FBS) was a product of PAN Biotech, Germany. 12-well Transwell inserts (0.4 μm pore size, 1.13 cm2 surface area), 6-, 12-, and 96-well plates all were purchased from Costar, Corning, USA. MTT assay dye and DMSO were purchased from Amresco, USA. Hoechst 33,258 was purchased from Solarbio, China. Chlorpromazine, β-cyclodextrin, Wortmannin and Amiloride were all purchased from Sigma-Aldrich, USA.
Hoechst 33258
Manufactured by Solarbio
Sourced in China, United States
Hoechst 33258 is a fluorescent dye used in biological research. It binds to DNA and emits blue fluorescence when excited by ultraviolet light. The dye can be used to stain and visualize DNA in various applications such as fluorescence microscopy and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 880, by Zeiss (10 mentions)
Fluorescence microscope, by Olympus (4 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
B27 supplement, by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by PAN Biotech (2 mentions)
Fluorescence microscope, by Nikon (2 mentions)
Ab53707, by Abcam (2 mentions)
Formic acid, by Solarbio (2 mentions)
Fluorescence microscope, by Zeiss (2 mentions)
Bx50 fla, by Olympus (2 mentions)
Infinite m200, by Tecan (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
3 3 diaminobenzidine (dab), by Gene Tech (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Amiloride, by Merck Group (1 mentions)
Chlorpromazine, by Merck Group (1 mentions)
Wortmannin, by Merck Group (1 mentions)
66 protocols using hoechst 33258
1
Evaluating Recombinant porcine IFN-α in Caco-2 Cells
2
Neurochemical Analysis of Cultured Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gibco Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), neurobasal medium and B27 supplement were purchased from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). Ketamine hydrochloride was purchased from Fujian Gutian Yuanhang Medical Company Ltd., Co. (Ningde, China). 17β-Estradiol, pregnenolone (PREG), methyltestosterone (MT), dansyl chloride, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Testosterone was provided by the National Institute for Control of Pharmaceutical and Biological Products (Beijing, China). Ethyl acetate-n-hexane was provided by Concord Technology Co., Ltd. (Tianjin, China). Trypsin and Hoechst 33258 were purchased from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). The in situ cell death detection kit was from Roche Applied Science (Mannheim, Germany).
3
Hoechst Staining for Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell apoptosis was determined by using Hoechst staining. In Hoechst staining assay, cells were fixed in 4% paraformaldehyde at room temperature for 30 min, and stained with Hoechst 33258 (Solarbio, Shanghai, China) for 10 min and observed under a fluorescence microscope (Nikon, Tokyo, Japan).
4
LoVo Cell Transfection and Hoechst Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The LoVo cells were cultured in 6-well plates. After 48 h of transfection with miR-518a-3p mimics, inhibitors or negative control, the cells were washed with PBS and then stained with Hoechst 33258 (10 μg/ml) (Beijing Solarbio Science & Technology Co., Ltd.) for 5 min before being washed 3 times with PBS.
5
Evaluating Apoptosis via Hoechst Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptotic cells were evaluated by morphologic observation using Hoechst 33,258 (Solarbio, Beijing, China) staining. Specifically, cells at logarithmic growth were seeded in 6-well plates by density of 1×105 per well. After transfected with siRNA for 2 days, cells were fixed with 4% paraformaldehyde for 20 min at room temperature, and subsequently stained with Hoechst 33,258 at 37°C for 30 min. Cells were observed with a fluorescence microscope (Nikon, Tokyo, Japan).
6
Hoechst-33258 Staining of Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis levels of H9c2 cells were detected using Hoechst-33258 staining (Solarbio Life Sciences, Beijing, China). Briefly, after drug treatment, cells were incubated with Hoechst-33258 at 37°C for 20 min and followed by two washes with PBS to remove free dye. Afterward, images were collected by using a fluorescence microscope.
7
Immunofluorescence Staining of FFPE Intestinal Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To prepare formalin-fixed and paraffin-embedded (FFPE ) tissue sections for immunofluorescence, samples from the duodenum, jejunum, and ileum were fixed in 4% paraformaldehyde and embedded in paraffin according to conventional methods. Briefly, 5-μm thick sections were cut onto gelatinized slides. The slides were deparaffinized and rehydrated and placed in an antigen retrieval solution (SolarBio, China) in a boiling water bath for 30 min. The slides were blocked using goat serum and incubated at room temperature for 20 min. Sections were then stained with mouse anti-chicken IgA-FITC (SouthernBiotech, USA) in humidified chambers overnight at 4°C. After washing with PBS, the nuclei were stained using Hoechst 33,258 (SolarBio, China). The autofluorescence of the FFPE tissue sections were diminished using sodium borohydride according to the method of Davis et al. (2014 (link)). A total of 162 sections (3 groups × 6 birds per group × 3 intestinal segments with 3 replicates) were observed using an inverted microscope (Axio Examiner ZEISS, Germany), and ZEN lite for Windows was used to photograph images under 100× magnification. It should be ensured that more villus and crypt appeared under the microscope; the number of IgA-positive cells of per villus was calculated by Image J software (NIH, USA).
8
Immunostaining of Psoriasis Skin Biopsies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Biopsies obtained from the lesional skin of psoriasis patients and the skin of healthy controls or from IMQ-induced mice were fixed with a 12% formaldehyde solution and embedded in paraffin. For immunofluorescence staining, cells or skin biopsy specimens were incubated with the following primary antibodies overnight at 4°C: anti-K17 (1:1000, ab53707, Abcam, UK), anti-ENO1 (1:400, ab227978, Abcam, UK), and anti-Ki67 (1:1000, ab15580, Abcam, UK). After three washes with PBS, the cells were incubated with Cy3- or FITC-conjugated secondary antibodies (1:200, BioLegend, San Diego, USA), and Hoechst 33258 (1:1000, Solarbio Technology, China) was applied to all cells to label the nuclei. Samples were analyzed using a confocal microscope (LSM880, Carl Zeiss, Germany). For immunohistochemical staining, the tissue sections were incubated with 0.3% H2O2 for 10 min and then incubated with one of the following primary antibodies: anti-K17 (1:1000, ab53707, Abcam, UK) or anti-ENO1 (1:400, ab227978, Abcam, UK) overnight at 4°C. Sections were subsequently incubated with an HRP-labeled goat anti-mouse/rabbit antibody (CWBIO, Peking, China) for 1 h at room temperature. DAB (Gene Tech, Shanghai, China) was used to detect biotinylated antibodies.
9
Bacterial Viability Assay with AlfB1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
P. damselae cells at the logarithmic growth stage were diluted to 105–106 CFU/ml in PBS. The bacteria were pre-incubated with rCtl24 (50 μg/ml) for 2 h or not, and then with 5 μM of AlfB1 for 2 h at 28°C. After three washes, bacteria were stained using 1 μg/ml of propidium iodide (PI) (C0080, Solarbio, Beijing, China) and Hoechst 33258 (B8030, Solarbio) for 15 min at room temperature. After staining, the bacteria were washed three times and suspended in PBS. For flow cytometry, the cell suspension was examined using a flow cytometer (BD FACSAria Fusion, BD, Franklin Lakes, NJ, USA). The data were analyzed using the FlowJo software (FlowJo LLC, Ashland, OR, USA). For confocal imaging, the cell suspension was examined in the multi-track mode using a confocal microscope (LSM 900 with Airyscan, ZEISS, Oberkochen, Germany). The images were analyzed and presented using the ZEN software program (Zeiss).
10
BSHX Granules and AGEs Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BSHX granules were purchased from Beijing Kangrentang Pharmaceutical Co. Ltd. (Beijing, China). The medicinal components of BSHX are listed in Table 1 . AGEs were purchased from Beijing Biosynthesis Biotechnology Co. Ltd. (Beijing, China) with a purity greater than 95% by high-performance liquid chromatography (HPLC). Antibodies against cleaved caspase-3, Bim, Histone H3, FoxO1, FoxO3a, α-Tubulin, and rabbit IgG were obtained from Cell Signaling Technology (Boston, MA, USA). 3-(4,5-Dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) was bought from Sigma-Aldrich Chemical Co. (Saint Louis, MO, USA). The Dulbecco's modified eagle's medium (DMEM) and fetal bovine serum (FBS) were purchased from PAN-Biotech GmbH (Aidenbach, Germany). The endothelial cell medium (ECM) was obtained from Beijing M&C Gene Technology Ltd. (Beijing, China). The lactate dehydrogenase (LDH) assay kit was purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Hoechst 33258 was obtained from Beijing Solarbio Science & Technology Co. Ltd. (Beijing, China). The enhanced BCA protein assay kit was obtained from Beijing TransGen Biotech Co. Ltd. (Beijing, China).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!