Alexa fluor 594

Manufactured by BD

Sourced in United States, United Kingdom

Alexa Fluor 594 is a fluorescent dye commonly used in biomedical research. It is characterized by its bright red emission and excellent photostability. The dye can be conjugated to a variety of biomolecules, including proteins, antibodies, and nucleic acids, to enable fluorescent labeling and detection.

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Lab products found in correlation

Alexa fluor 488, by BD (2 mentions) S500 statspin thermobrite slide hybridization denaturation system, by Abbott (1 mentions) Vysis centromere probe cep8 spectrum orange, by Abbott (1 mentions) Alexa flour 488, by BD (1 mentions) Cm1850 microtome, by Leica (1 mentions) M30 cytodeath, by Roche (1 mentions) In situ cell death detection kit, by Roche (1 mentions) Image pro software, by Media Cybernetics (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Hsp60, by Abcam (1 mentions)

Close spelling variants of this product

Alexa Fluor™ 594 Phalloidin (2 citations) Alexa Fluor 594-conjugated donkey anti-rat secondary antibody (2 citations)

5 protocols using alexa fluor 594

Identifying CTCs with Fluorescent Markers

Cited 1 time

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The experimental procedure and CTC identification were conducted as previous described [6 (link)]. Experiment was performed according to the product manufacture's instruction (Cytelligen). Briefly, samples on the coated CTC slides were subjected to Vysis Centromere Probe (CEP8) SpectrumOrange (Abbott Laboratories, Abbott Park, IL, USA) hybridization for 90 min using a S500 StatSpin ThermoBrite Slide Hybridization/Denaturation System (Abbott Molecular, Des Plaines, IL, USA), followed by incubation with Alexa Fluor 594 conjugated monoclonal anti-CD45 and Alexa Fluor 488 conjugated with monoclonal anti-GFAP (BD) as described above. Images of the identified tumor cells were collected using a fluorescence microscope (OLYMPUS, BX-53) equipped with a filter set (Omega Optical, Brattleboro, VT, USA) for DAPI (Cytelligen), Alexa Flour 488 (BD), Alexa Flour 594 (Cytelligen), and Spectrum Orange, TRITC (Vysis). CTC is defined as DAPI+, CD45-, and polyploidy CEP8 signal with or without visible GFAP signal.

Gao F., Cui Y., Jiang H., Sui D., Wang Y., Jiang Z., Zhao J, & Lin S. (2016). Circulating tumor cell is a common property of brain glioma and promotes the monitoring system. Oncotarget, 7(44), 71330-71340.

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Aortic Cryosectioning and Immunostaining

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Immediately after aortic stiffness measurements, mice were euthanized by carbon dioxide and isolated aortas were flash frozen in optimum cutting temperature freezing medium on dry ice. Aortas were sliced using a CM 1850 microtome (Leica Biosystems) to produce 10μm thick sections. Cryosections were fixed in cold 4% paraformaldehyde in PBS for 10 minutes, washed with PBS, and incubated with 0.3% Triton in PBS for 10 minutes. Subsequently, sections were blocked with 5% BSA in PBS for one hour and incubated with rat anti-mouse Me1 antibody conjugated with Alexa Fluor 594 (PharMingen) overnight at 4^oC. After washing with PBS, sections were incubated with Hoechst blue nuclear dye (Molecular Probes) for 1 hour at room temperature in the dark. At least 5 consecutive sections were examined for each aorta. Fluorescent signal was imaged on the Zeiss LSM710 Confocal AxioObserver Inverted Automated Microscope and quantified by Image J software (NIH).

Zhu W., Wang H., Wei J., Sartor G.C., Bao M.M., Pierce C.T., Wahlestedt C.R., Dykxhoorn D.M, & Dong C. (2018). Cocaine Exposure Increases Blood Pressure and Aortic Stiffness via the miR-30c-5p—Me1—ROS Pathway. Hypertension (Dallas, Tex. : 1979), 71(4), 752-760.

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Immunostaining of Drosophila Tissues

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Tissues were fixed in 3% paraformaldehyde in PBS for 30 min then immunostained as described previously [20 (link)]. Mouse anti-Coracle and anti-Armadillo antibodies (Developmental Studies Hybridoma Bank (DSHB)) and anti-Fibrillarin (Thermo Fisher Scientific) were used at 1:10, 1:400, and 1:400 dilution respectively. Goat anti-mouse IgG labeled with Alexa Fluor 594 (BD Biosciences) was used at 1:500 dilution as the secondary antibody. Guinea pig anti-Lov antibody, prepared and characterized as described previously [20 (link)], was used at a 1:50 dilution followed by goat anti-guinea pig IgG labeled with Alexa Fluor 594 or 488. DNA was stained with DAPI (4’,6-diamidino-2-phenylindole) at 1μg/ml prior to mounting in 70% glycerol. Fat body was simultaneously fixed and stained in 37% formaldehyde containing five units/ml of Oregon-Green 488nm Phalloidin (Molecular Probes) and DAPI at 1μg/ml.

Zhou F., Green S.R., Tsay M., Hsu S., Dibbs R, & Beckingham K.M. (2020). The roles of jim lovell and uninflatable in different endopolyploid larval tissues of Drosophila melanogaster. PLoS ONE, 15(8), e0237662.

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Tissue Histology and Fluorescence Analysis

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For histological, immunofluorescent and direct fluorescent analyses, tissue samples (eg. small intestines, tumors) were embedded in the OCT compound (Tissue-Tek), and cryo-sectioned (7 µm). Hematoxylin and Eosin staining was done on ethanol/acetic acid fixed slides. Apoptosis incidence was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) using the In Situ Cell Death Detection Kit (Roche Applied Science) and 4',6-diamidino-2-phenylindole (DAPI) counterstain. Quantitative analysis was done using ImagePro software (Media Cybernetics, Rockville, MD) on at least 6 different and independent microscopic fields for each treatment condition. Indirect immunofluorescence was performed by incubation with primary antibodies for cleaved PARP (Asp214) Human Specific (Cell Signaling), M30CytoDEATH (Roche Applied Science), or Ki-67 (abcam) for 1 h at room temperature or overnight at 4°C followed by secondary antibodies labeled with either Alexafluor⁴⁸⁸ or Alexafluor⁵⁹⁴ (BD Biosciences). Sections were counterstained with DAPI and slides were mounted using anti-fade Fluorosafe reagent (Calbiochem).

Alexeev V., Lash E., Aguillard A., Corsini L., Bitterman A., Ward K., Dicker A.P., Linnenbach A, & Rodeck U. (2014). Radiation protection of the gastrointestinal tract and growth inhibition of prostate cancer xenografts by a single compound. Molecular cancer therapeutics, 13(12), 2968-2977.

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Profiling Platinum Sensitive and Resistant Ovarian Cancer

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Intra-patient paired platinum sensitive and resistant high grade serous ovarian carcinoma cell lines (PEA1 vs PEA2 and PEO14 vs PEO23 respectively) were obtained from Dr. Simon Langdon (Edinburgh, UK) [29 (link)]. The cell lines PEA1 and PEO14) were derived prior to the onset of platinum resistance, whereas PEA2 and PEO23 were derived following acquired clinical platinum resistance and were cultured and maintained in RPMI-1640 medium, supplemented with 10% foetal calf serum, 1% Penicillin Streptomycin and L-Glutamine (all GIBCO, UK) at 37°C/5% CO₂.
Antibodies used were IL-8-RA/CXCR1 and IL-8-RB/CXCR2 (R&D systems, UK); Alexa Fluor 594 (BD Diagnostics, UK); Bcl-2 and p-Bad S136 (Cell Signalling Technology, UK); HSP60 (Abcam, UK); goat anti-mouse and goat anti-rabbit HRP conjugated secondary antibodies (DAKO, Denmark). Blocking of IL-8-RA/B Signalling was achieved using a small molecule antagonist SCH563705 [30 (link), 31 (link)].

Stronach E.A., Cunnea P., Turner C., Guney T., Aiyappa R., Jeyapalan S., de Sousa C.H., Browne A., Magdy N., Studd J.B., Sriraksa R., Gabra H, & El-Bahrawy M. (2015). The role of interleukin-8 (IL-8) and IL-8 receptors in platinum response in high grade serous ovarian carcinoma. Oncotarget, 6(31), 31593-31603.

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