Silica nanoparticles (SiNPs, 637246) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and Turbofect transfection reagent were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Annexin V-PE/7-AAD kit, total protein extraction kit, and BCA protein assay kit were purchased from Nanjing Keygen Biotech Co., Ltd. (Nanjing, Jiangsu, China). DAPI (4′, 6-diamidino-2-phenylindole) was purchased from Beyotime Institute of Biotechnology (Shanghai, China). Anti-GRP78 antibody (C50B12), anti-cleaved Caspase-3 antibody (9664), and anti-mouse and anti-rabbit HRP-linked secondary antibodies (7076P2, 7074P2) were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-ERO1α antibody (H00030001-M01) was purchased from Abnova (Taipei, Taiwan). Anti-BCL-2 antibody (ab182858), anti-BCL-xL antibody (ab32370), anti-MCL-1 antibody (ab32087), anti-PUMA antibody (ab33906), anti-BAX antibody (ab32503), anti-BAD antibody (ab32445), and anti-mouse and anti-rabbit fluorescent secondary antibodies (ab150077, ab150115) were purchased from Abcam Ltd. (Cambridge, MA, USA). Anti-CHOP antibody (sc-7351) was purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Anti-β-actin antibody (KM9001T) was purchased from Tianjin Sungene Biotechnology Inc. (Tianjin, China).
Dapi 4 6 diamidino 2 phenylindole
Manufactured by Beyotime
Sourced in China
DAPI (4′,6-diamidino-2-phenylindole) is a fluorescent dye commonly used in biological research. Its core function is to bind to adenine-thymine (A-T) rich regions of DNA, emitting a blue fluorescence when exposed to ultraviolet (UV) or violet light. This property makes DAPI a useful tool for DNA staining and visualization in various applications, such as fluorescence microscopy and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescence microscope, by Leica camera (3 mentions)
Fish kit, by RiboBio (2 mentions)
Fv10 laser scanning confocal microscope, by Olympus (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Bca protein assay kit, by Beyotime (2 mentions)
Pan cadherin c1821, by Merck Group (2 mentions)
Fish kit, by GenePharma (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Multiskan fc, by Thermo Fisher Scientific (2 mentions)
Fluorescence microscope, by Nikon (2 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Turbofect transfection reagent, by Thermo Fisher Scientific (1 mentions)
Annexin 5 pe 7 aad kit, by Keygen Biotech (1 mentions)
Total protein extraction kit, by Keygen Biotech (1 mentions)
Bca protein assay kit, by Keygen Biotech (1 mentions)
Rapamycin sirolimus, by Selleck Chemicals (1 mentions)
Trem2, by Abcam (1 mentions)
Alexa fluor 790, by Abcam (1 mentions)
Rpmi 1640, by Sartorius (1 mentions)
41 protocols using dapi 4 6 diamidino 2 phenylindole
1
Silica Nanoparticles-induced ER Stress Pathway
2
Investigating Autophagy and Inflammation Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ABT-263 (Navitoclax), 3-MA (3-methyladenine) and Rapamycin (Sirolimus) were purchased from Selleck Chemicals (TX, USA). The primary Rabbit monoclonal antibodies of Bcl-2, Bax, Beclin-1(Becn), Atg5, LC-3, Trem-2, and Tubulin-α conjugated with Alexa Fluor® 790 were acquired from Abcam (Cambridge, UK). The secondary antibodies Goat anti-Rabbit IgG H&L (Alexa Fluor®680) was purchased from Abcam as well. The DAPI (4′, 6-diamidino-2- phenylindole) was obtained from Beyotime (Shanghai, China). The F4/80-PE anti-mouse antibody, PE Rat IgG2a isotype and biotin anti-mouse CD16/32 antibody were purchased from Biolegend (San Diego, CA). The Phallodin-iFluor 633 Conjugate was purchased from ATT Bioquest® (Sunnyvale, CA). The cell culture reagents, RPMI 1640 and FBS (fetal bovine serum) were purchased from Biological Industries (Cromwell, CT). The RT2 First Strand kit and Profiler PCR Array (PAXX-084) and SYBR Green qPCR master mix were purchased from Qiagen (Hilden, Germany). Proteome Profiler Array of Mouse cytokine (ARY006) was purchased from R&D systems, Inc. (MN, USA).
3
Evaluating Compound RRx-001 in Cell and Animal Models
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RRx-001 was purchased from MedChemExpress (Monmouth Junction, NJ, United States) and dissolved in dimethyl sulfoxide (DMSO) for cell experiments, and dissolved in 10% DMSO and 90% corn oil for animal experiments. Lipopolysaccharide (LPS) and ATP were purchased from Sigma (St. Louis, United States) and dissolved in PBS for cell experiments, and dissolved in normal saline for animal experiments. DCFH-DA (2′, 7′-dichlorodihydrofluorescein diacetate) was purchased from Solarbio (Beijing, China). Propidium iodide (PI), Hoechst 33,342, and DAPI (4′, 6-diamidino-2-phenylindole) were purchased from Beyotime Biotechnology (Shanghai, China).
4
Quantifying Cellular Oxidative Stress
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Detection of ROS was performed on frozen sections using dihydroethidium (D7008, Sigma-Aldrich, United States) which was diluted as 1:500 for 30 min at 37°C in a dark incubator. DAPI (4′, 6-diamidino-2-phenylindole, Beyotime, China) was added after the slides were dry and incubated for 10 min in the dark at room temperature. Slides were washed three times for 5 min each in phosphate buffered saline (pH 7.4) on a shaker. After the plate was blocked with an antifade mounting medium (Servicebio, China), the images were observed and acquired under a fluorescence microscope.
5
Immunohistochemical Analysis of Aquaporin-4 and GFAP Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After being deparaffinized and rehydrated, the slices were antigen-retrieved in antigen retrieval buffer and blocked with goat serum. Then, the slices were incubated with AQP4 antibody (rabbit anti-AQP4 antibody; 1: 50 diluted in PBS; Proteintech) and glial fibrillary acidic protein (GFAP) antibody (mouse anti-GFAP antibody; 1: 50 diluted in PBS; Santa Cruz, Dallas, TX, USA) overnight at 4°C. After being rinsed in PBS, the slices were incubated with corresponding secondary antibodies (Cy3-conjugated goat anti-rabbit IgG, FITC-conjugated goat anti-mouse IgG; 1: 200 diluted in PBS; Beyotime) for 90 min at room temperature in the dark. Thereafter, the slices were rinsed in PBS and incubated with DAPI (4′,6-diamidino-2-phenylindole) (Beyotime) for nuclear counterstaining. The slices were observed using a fluorescence microscope (OLYMPUS; model: BX53) with a 400× magnification.
6
TUNEL Assay for Apoptosis Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Terminal Deoxynucleotidyl Transferase dUTP Nick End Labeling (TUNEL) staining was conducted using the One-step TUNEL Apoptosis in situ Assay Kit (KeyGen BioTECH, Nanjing, CHN) according to the manufacturer’s instructions. Cells with multiple DNA breaks were labeled with tetramethylrhodamine, and the nuclei were stained with DAPI (4′,6-diamidino-2-phenylindole) (Beyotime Biotechnology). Cell apoptosis was analyzed using New Dual-CCD DP80 Microscope Digital Camera (Olympus, Tokyo, Japan).
7
LINC01087 RNA Expression in Glioblastoma Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The LINC01087 probe was designed and synthesized by RiboBio (Guangzhou, China), and the sequence could be provided upon request. In view of the instructions, the probe signal was tested by FISH kit (RiboBio, Guangzhou, China). In short, GBM cells were immobilized in 4% formalin for 15 min. Cells were prehybridizated in PBS and then hybridized in hybridization solution at 37°C for 30 min. Next, the nuclei were re-stained by DAPI (4′, 6-diamidino-2-phenylindole) staining (Beyotime, China). Images were caught by a fluorescence microscope (Leica, Germany).
8
Docetaxel-resistant Breast Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human breast cancer cell lines MDA-MB-231, docetaxel (TXT)-resistant SKBR3, TXT-resistant BT474, TXT-resistant MCF7, and TXT-resistant MDA-MB-231 were obtained from Dr. Wang HH. All of these cell lines were cultured according to the ATCC instructions. siRNA and Lipofectamine™ 2000 were purchased from Thermo Scientific Dharmacon®. SYBR Green Real-time PCR Master Mix was purchased from Toyobo (Osaka, Japan). RevertAid™ First Strand cDNA Synthesis kit was obtained from MBI (Fermentas, Hanover, MD, USA). Docetaxel (Aventis Pharmaceuticals, Bridgewater, NJ, USA) was stored at a concentration of 10 mg/mL (12.6 mM) in 13% w/w ethanol at 4 °C and diluted in medium before use. Cell Counting Kit-8 was obtained from Roche. TUNEL assay was performed by use of In Situ Cell Death Detection Kit (Fluorescein, Roche, Switzerland). DAPI (4,6-diamidino-2-phenylindole) was purchased from Beyotime (Shanghai, China). Hoechst 33342 was purchased from Sigma-Aldrich China (Shanghai, China).
9
Immunofluorescence Staining of Chondrocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Following overnight culture in a 24-well plate, cells were fixed with 4% paraformaldehyde for 10 min and then treated with 0.2% Triton X-100 for 15 min. After blocking with 5% goat serum for 30 min, cells were incubated overnight at 4°C in a refrigerator with primary antibodies against type II collagen and SOX-9 primary (1:100). The following morning, cells were incubated with Alexa Fluor-555 labeled secondary antibody (Beyotime Institute of Biotechnology) for 2 h at room temperature, stained for 5 min at room temperature with DAPI (4′,6-diamidino-2-phenylindole) (Beyotime Institute of Biotechnology) and finally observed under a fluorescence microscope (Olympus Corporation, Tokyo, Japan).
10
Immunostaining of Osteogenic Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BMSCs were fixed with 4% paraformaldehyde after 7 days of growth on the CPC surface. Cells were permeabilized with 0.5% Triton X-100 for 10 min. Samples were washed thrice with PBS for 5 min and incubated with 3% bovine serum albumin solution for 30 min, followed by incubating with primary mouse anti-osteopontin (OC, 1:500, Abcam, UK) and rabbit anti-RUNX2 (1:500, Affinity, China) antibodies overnight at 4 °C. After washing thrice for 5 min each in PBS, Alexa Fluor 594-labeled donkey anti-mouse and Alexa Fluor 488-labeled donkey anti-rabbit (1:1000, Abcam, UK) were mixed and incubated for 2 h at 37 °C in dark. Samples were then washed with PBS and nuclei were stained with 10 ng/mL DAPI (4′,6-diamidino-2-phenylindole, Beyotime, China). Finally, the samples were imaged by a confocal laser scanning microscope (Zeiss LSM 800, Germany).
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!