Acridine orange

Manufactured by ImmunoChemistry Technologies

Sourced in United States

Acridine orange is a fluorescent dye used for staining nucleic acids in biological samples. It can bind to both DNA and RNA, emitting different colored fluorescence depending on the type of nucleic acid. This property makes acridine orange a useful tool for various applications in cell biology and microscopy.

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Lab products found in correlation

Lsm800 confocal microscope, by Zeiss (2 mentions) Lsm 880, by Zeiss (2 mentions) Lamp1, by Cell Signaling Technology (1 mentions) Egf receptor, by Cell Signaling Technology (1 mentions) Lysotracker red dnd 99, by Thermo Fisher Scientific (1 mentions) α tubulin, by Merck Group (1 mentions) Phospho s6 s235 236, by Cell Signaling Technology (1 mentions) Ca 074, by Apexbio (1 mentions) Parp1, by Cell Signaling Technology (1 mentions) Lamin a c, by Cell Signaling Technology (1 mentions) Bafilomycin a1, by Merck Group (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Anti flag m2 affinity gel, by Merck Group (1 mentions) β actin, by Merck Group (1 mentions) Zen 3, by Zeiss (1 mentions) Hoechst 33342, by ImmunoChemistry Technologies (1 mentions) Clathrin, by Abcam (1 mentions) Grp78, by Cell Signaling Technology (1 mentions) Celltrace violet, by Thermo Fisher Scientific (1 mentions) Vinculin, by Merck Group (1 mentions)

Spelling variants of this product

Acridine Orange (AO, (4 citations)

4 protocols using acridine orange

Curcumin-probe mediated autophagy analysis

Cited 1 time

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The chemicals used in our experiments were: LysoTracker Red DND-99 (Invitrogen, L7528), Magic RedTM cathepsin B reagent with acridine orange (Immunochemistry Technologies, LLC, 937/6130), CA-074 (APExBIO, A1926), bafilomycin A1 (Baf, Sigma, B1793), acridine orange (AO) (Immunochemistry Technologies, LLC, 6130) and [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS) (Promega, G5421). The Curcumin-probe (Curcumin-P) was readily synthesized by mono-alkylation of curcumin by propargyl bromide and its structure was verified by H-NMR, C-NMR and high resolution mass spectrometry [40 (link)].
The antibodies used in our experimentsincluded: microtubule-associated protein 1 light chain 3 (LC3) (Sigma, L7543), p62 (Sigma, P0067), Tsc2 (Cell Signaling Technology, 4308), phospho-S6(S235/236) (Cell Signaling Technology, 2211), S6 (Cell Signaling Technology, 2317), β-actin (Sigma, A5441), α-tubulin (Sigma, T6199), Lamin AC (Cell Signaling Technology, 2032), EGF receptor (Cell Signaling Technology, 4267), LAMP1 (Cell Signaling Technology, 9091), PARP1 (Cell Signaling Technology, 9542), TFEB (Bethyl Laboratories, A303-673A), FLAG (Sigma, F1804), 14-3-3 (Cell Signaling Technology, 9638), ANTI-FLAG® M2 Affinity Gel (Sigma, A2220).

Zhang J., Wang J., Xu J., Lu Y., Jiang J., Wang L., Shen H.M, & Xia D. (2016). Curcumin targets the TFEB-lysosome pathway for induction of autophagy. Oncotarget, 7(46), 75659-75671.

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Lysosomes in RANKL-primed BMDMs

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Lysosomes in RANKL‐primed primary BMDMs treated with PGDHC were evaluated using Acridine Orange (Immunochemistry Technologies, Cat# 6130) at a 1:1000 dilution and Hoechst 33342 (Immunochemistry Technologies, Cat# 639) nuclear counterstain at 0.5% v/v concentration, according to the manufacturer's recommendation. The images were acquired using a Zeiss LSM 800 confocal microscope and analyzed using the Zen (Black edition), Zen 3.2 (Blue edition) and Image J software.

Duarte C., Yamada C., Garcia C., Akkaoui J., Ho A., Nichols F, & Movila A. (2022). Crosstalk between dihydroceramides produced by Porphyromonas gingivalis and host lysosomal cathepsin B in the promotion of osteoclastogenesis. Journal of Cellular and Molecular Medicine, 26(10), 2841-2851.

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Multiparametric Analysis of Cell Cultures

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Lysotracker, CellTrace Violet, FM lipophilic styryl (FM 1-43FX) and SYTO RNASelect dyes were from Life Technologies (Thermo Fisher Scientific). Acridine Orange was from ImmunoChemistry Technologies (Bloomington, MN, USA). PropidiumIodure (PI) and Annexin V were from BD Bioscience (FITC-Annexin V Apoptosis Detection Kit). Bafilomycin A1 (BafA1; sc-201550) was from Santa Cruz Biotechnologies whereas NH4Cl (254134) was from Sigma Aldrich.
The following primary antibodies were used for immunofluorescence, immunoblotting (IB) and flow cytometry (FACS) assays: V-ATPase G1 (16143-1-AP, Proteintech), Vinculin (V9131, Sigma Aldrich), Tsg101 (14497-1-AP, Proteintech), Ago2 (10686-1-AP, Proteintech), Clathrin (ab23440, Abcam), CD63 (sc-15363, Santa Cruz), CD9 (IB: 10626D, Thermo Fisher; FACS: 130-103-988, Miltenyi), CD81 (130-107-982, Miltenyi), Calnexin (ab31290, Abcam), Nestin (MAB 1259, R&D), Tuji (T3952, Sigma Aldrich), GFAP (G9269, Sigma Aldrich), CD11b (20991-1-AP, Proteintech), O4 (O7139, Sigma Aldrich), Olig2 (AV32753, Sigma Aldrich), CD31 (ab28364, Abcam), Vimentin (130-106-369, Miltenyi), POU3F2 (12137, Cell Signaling), HOXA10 (TA590263, Origen), HOXA7 (ab211521, Abcam), V-ATPase G2 (25316-1-AP, Proteintech), ARF6 (6ARF01, ThermoFisher Scientific), and GRP78 (3177, Cell Signaling).

Bertolini I., Terrasi A., Martelli C., Gaudioso G., Di Cristofori A., Storaci A.M., Formica M., Braidotti P., Todoerti K., Ferrero S., Caroli M., Ottobrini L., Vaccari T, & Vaira V. (2019). A GBM-like V-ATPase signature directs cell-cell tumor signaling and reprogramming via large oncosomes. EBioMedicine, 41, 225-235.

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Detecting Autophagy via Acridine Orange

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To determine the role of autophagy in IM-induced cell death, acridine orange (ImmunoChemistry Technologies, Davis, CA, USA) was used to determine if the drug increased acidic properties within the cell (acidic organelles), indicating the presence of increased lysosome content. Cells were plated into a 12-well plate and allowed to reach confluency. Following this, 5 µM acridine orange was added to medium for 30 min. Cells were then washed once with PBS and imaged using confocal microscopy (LSM880, Zeiss, Oberkochen, Germany). The number of positive cells were counted using Image J (National Institutes of Health, Bethesda, MD, USA).

Walmsley R., Steele D.S., Ellison-Hughes G.M., Papaspyros S, & Smith A.J. (2022). Imatinib Mesylate Induces Necroptotic Cell Death and Impairs Autophagic Flux in Human Cardiac Progenitor Cells. International Journal of Molecular Sciences, 23(19), 11812.

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