Dulbecco's modified Eagle's medium (DMEM) was from Gibco (USA). Fetal bovine serum (FBS) was from Hyclone Lab Inc. (USA). Sulforhodamine B (SRB), Hoechst 33258, and 2′,7′-dichlorodihydrofluorescein (DCHF) were from Sigma Co. (USA). Acridine orange was from Amresco (USA). Primary antibodies against β-actin and secondary antibody (horseradish peroxidase) were from Santa Cruz Biotechnology (USA). Primary antibody against procaspase 3 was from Cell Signaling Technology (USA). All other reagents were ultrapure grade.
Acridine orange
Manufactured by Avantor
Sourced in United States, United Kingdom
Acridine orange is a fluorescent dye commonly used in microscopy and cell biology applications. It can selectively stain nucleic acids, allowing for the visualization of cellular structures such as DNA and RNA. The dye exhibits different fluorescent properties when bound to double-stranded DNA versus single-stranded DNA or RNA.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Sulforhodamine b srb, by Merck Group (2 mentions)
Hoechst 33258, by Merck Group (2 mentions)
Alexa fluor 488 donkey anti rabbit igg, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Propidium iodide, by Merck Group (2 mentions)
β actin, by Santa Cruz Biotechnology (2 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Propidium iodide, by US Biological (1 mentions)
Dulbecco s modified eagle s medium dmem, by Caisson (1 mentions)
Collagen type 1, by US Biological (1 mentions)
Lc3b polyclonal antibody, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Caisson (1 mentions)
Triton x 100, by US Biological (1 mentions)
Nf κb p65, by Santa Cruz Biotechnology (1 mentions)
L α phosphatidylcholine, by Merck Group (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Accuri c6, by BD (1 mentions)
11 protocols using acridine orange
1
Cell Proliferation and Apoptosis Assay
2
Apoptosis and Autophagy Assay Protocol
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Ferric chloride hexahydrate (FeCl3.6H2O) was purchased from Fisher Chemicals and tannic acid (C76H52O46) was from Loba Chemie; Dulbecco’s modified Eagle’s medium (DMEM), Dulbecco’s modified Eagle’s medium Ham’s F-12 (DMEM/F-12), and Trypsin-EDTA were purchased from Caisson Laboratories. Fetal bovine serum (FBS) was from HyCloneTM Thermo Fisher Scientific. Collagen type I, ribonuclease A (Rnase A), triton X-100, propidium iodide (PI), and annexin V (FITC) apoptosis detection BioAssay™ kit were purchased from US Biological. Sodium azide (NaN3) and potassium thiocyanate (KSCN) were procured from BDH Laboratory. Dimethyl sulfoxide (DMSO) and monodensylcadavarine (MDC) were purchased from Sigma-Aldrich; nitric acid (HNO3) and ethanol were procured from ACL Labscan; and formalin was purchased from Gammaco. 3-dimethylthiazol-2,5-diphenyltetrazolium bromide (MTT) and acrylamide/bis-acrylaminde were purchased from Bio Basic Inc. and etramethylethylenediamine (TEMED) was procured from Invitrogen. Hydrochloric acid (HCl) was purchased from J.T. Baker; methanol was procured from Northern Chemicals and Glasswares Ltd.; and acridine orange was purchased from AMRESCO. LC3B polyclonal antibody and goat anti-rabbit lgG (H + L) secondary antibody, DyLight 488, were purchased from Thermo Fisher Scientific.
3
Oxidized LDL-Induced Cellular Responses
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DMEM was from Gibco (USA). Fetal bovine serum (FBS) was from Hyclone Lab Inc. (USA). L-α-phosphatidylcholine, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 2′,7′-dichlorodihydrofluorescein diacetate (DCFH), and JC-1 were from Sigma Co. (USA). Acridine orange was from Amresco (USA). Ox-LDL was from Beijing Union-Biology Co. (China). Primary antibodies against ANXA7, NF-κB p65, GAPDH, and secondary antibody (horseradish peroxidase) were from Santa Cruz Biotechnology (USA). Secondary antibody for immunofluorescence, donkey anti-rabbit IgG Alexa Fluor-488, was purchased from Life Technologies (USA). All other reagents were of ultrapure grade.
4
Autophagy Detection in U2OS Cells
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For the detection of autophagy, U2OS cells were treated with Polyphyllin VI or 0.1% DMSO for 24 hrs. Following treatment, the cells were stained with 1 μg/mL acridine orange (Amresco, Solon, OH, USA) in PBS for 30 mins at 37°C. Cells were then observed using the BD Accuri C6 flow cytometry.
5
Quantifying Osteoclast Formation
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BMMs were cultured with M-CSF and RANKL for 5 days prior to incubation in 0.01% acridine orange (Amresco) for 15 min at room temperature, a PBS wash and staining in 0.1 mol/L Cacl2 for 2 min. Fluorescence images were obtained using a fluorescence microscope (Leica Microsystems, Mannheim, Germany; 490 nm excitation filter and 525 nm arrest filter).
6
Combination Therapy for Breast Cancer
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The oils of Borage and Medium Chain Triglyceride (MCT) were obtained from the electronic store iHerp.com. Tween 80 and Span 20 were purchased from Al Shafei medical and Scientific Equipment, Est (Jeddah, KSA). Docetaxel (DTX) was kindly provided by King Abdulaziz University Hospital. Thymoquinone (TQ) and Sulforhodamine B (SRB) dye were obtained from Sigma-Aldrich (USA). The Coomassie blue dye was purchased from Cayman Chemical (Michigan, US). Annexin V-FITC apoptosis detection kit was obtained from Invitrogen Abcam (UK). Acridine Orange and rapamycin were obtained from BDH Chemicals Ltd (England, UK) and Enzo Life Sciences (Lausen, Switzerland), respectively. Human Breast cancer cell lines, MCF-7 and MDA-MB-231, were obtained from ATCC (American Type Tissue Culture Collection, Manassas, VA, USA).
7
Acridine Orange Sperm DNA Integrity Assay
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Acridine orange (AO, 3, 6-bis (dimethylamino) acridine, hemi (zinc chloride) salt, BDH Chemicals Ltd., Poole, England) treatment assays are used to assess the sperm DNA vulnerability to acid-induced denaturation in situ by quantifying the metachromatic shift of AO fluorescence from green (double-stranded DNA) to red (denatured DNA). The AO test was carried out as described by Moretti et al. [19 (link)]. The slides were observed and scored with a Leitz Aristoplan fluorescence Microscope (Leica, Wetzlar, Germany) equipped with a 490 nm excitation light and 530 nm barrier filter. Three hundred sperm nuclei for each sample were analyzed and classified as green or red (sometimes, orange yellow) depending on fluorescence. The sperm heads that exhibited a green stain had double-stranded DNA; the sperm heads showing a spectrum of yellow orange to red fluorescence had single-stranded DNA. The results were expressed as the percentage of spermatozoa that showed normal DNA (green fluorescence).
8
Acridine Orange Staining for Acidic Vesicular Organelles
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To detect the development of acidic vesicular organelles (AVOs), staining with acridine orange was applied according to published procedures with modifications.18 (link) Following treatments for 48 hours, the positive control cells were exposed to the autophagy-inducing factor rapamycin (10 μg/well) (Enzo Life Sciences, Lausen, Switzerland).19 (link) After washing, the collected cells were stained with 1 μg/mL acridine orange (BDH Chemicals Ltd., Poole, England) for 30 minutes in the dark at 37°C. The samples were evaluated via BD FACSAria II Flow Cytometer (BD Biosciences, San Jose, CA, USA) and quantified by FACS DiVa software version 6.1.3.
9
Comprehensive Cell Viability Assay
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RPMI-1640 medium (Gibco, USA), fetal bovine serum(FBS, Gibco, USA), MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, Fluka, USA), acridine orange (AO, Amresco, USA), ethidium bromide (EB, Sigma, USA), Wright-Giemsa dye solution (Nanjing Jiangcheng Bioengineering Institute, Nanjing, China), RNase A (Fermantas, Canada), proteinase K (Fermantas, Canada), ECL plus kit (Beyotime Institute of Biotechnology, China), DL 2000 DNA Marker (Takata, Japan), prestained protein marker (fermentas, Thermo Scientific, USA), antibody to GAPDH, GRP78, GADD153, Bcl-2, Bax, p53, survivin, Actived-Caspase 3 p17 (Bioworld Technology, MN, USA), horseradish peroxidase (HRP)-conjugated secondary antibody (MultiSciences, China). All other biochemicals and chemicals used in the experiment were of analytical grade.
10
Apoptosis Evaluation in Cell Lines
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DMEM was obtained from Gibco BRL Co. (USA). Fetal bovine serum was from Hyclone Laboratories Inc. (USA). Acridine orange (AO) was from Amresco (USA). Hoechst 33258, 2′,7′-dichlorodihydrofluorescin (DCHF), JC-1, propidium iodide (PI), and sulforhodamine B (SRB) were from Sigma Co. (USA). The antibodies against p53, ANXA7, NF-κB p65, β-actin, and horseradish peroxidase-conjugated secondary antibodies were from Santa Cruz Biotechnology (USA). Secondary antibodies for immunofluorescence, donkey anti-rabbit IgG Alexa Fluor-488, were purchased from Life Technologies (USA). All the other reagents were of ultrapure grade.
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