Cy3 and cy5

Manufactured by Jackson ImmunoResearch

Sourced in Sweden

Cy3 and Cy5 are fluorescent dyes commonly used in molecular biology and immunoassay techniques. Cy3 emits light in the orange-red region of the visible spectrum, while Cy5 emits light in the far-red region. These dyes are often used for labeling proteins, nucleic acids, and other biomolecules to facilitate detection and visualization in various applications.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Lsm 700, by Zeiss (1 mentions) Mouse anti gfp, by Roche (1 mentions) Dcp 1, by Cell Signaling Technology (1 mentions) Lysotracker, by Thermo Fisher Scientific (1 mentions) Axio observer microscope, by Zeiss (1 mentions) In situ cell death detection kit, by Roche (1 mentions) Alexa fluor 568, by Thermo Fisher Scientific (1 mentions) Fluorescein avidin d, by Vector Laboratories (1 mentions) Sp8 laser scanning confocal microscope, by Leica (1 mentions) Triton x 100, by Merck Group (1 mentions)

Close spelling variants of this product

Cy3- and Cy5-conjugated secondary antibodies Cy3 and Cy5 conjugated goat α-HRP Cy3/Cy5

3 protocols using cy3 and cy5

Immunohistochemistry Embryo Staining

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For immunohistochemistry embryos were fixed and stained according to standard procedures. Guinea pig anti-SIMU (30 (link)) and guinea pig anti-Drpr (32 (link)) were used at a 1:5000 and 1:100 concentrations, respectively. Rabbit anti-activated caspase 3 (Dcp-1) (Cell Signaling) and mouse anti-GFP (Roche) were used at 1:100 concentration. Rabbit anti-Crq antibody (1:500) is a gift from N. Franc. Rabbit anti-Srp antibody (1:100) is a gift from J. Casanova, K. Campbell and N. Martin. Rabbit anti-Peroxidasin antibody (1:2000) is a gift from Jiwon Shim. Fluorescent secondary antibodies (Cy3/and Cy5/Jackson ImmunoResearch; Alexa Fluor 488/Molecular Probes) were used at 1:200 dilutions. For TUNEL labeling embryos were re-fixed, washed and labeled with the In Situ Cell Death Detection kit (Roche) according to the manufacture instructions. Images were acquired on a confocal microscope Zeiss LSM 700 or on a Zeiss Axio Observer microscope equipped with an Apotome system using the AxioVision software. 75% Glycerol solution was used as the imaging medium.
Live imaging was carried out by dechorionating embryos (stage 15), mounting them under Halocarbon oil, injecting 2–3% egg volume of LysoTracker (Molecular Probes) as described in Ref. (33 (link)). Recording started 30 min following injection.

Shlyakhover E., Shklyar B., Hakim-Mishnaevski K., Levy-Adam F, & Kurant E. (2018). Drosophila GATA Factor Serpent Establishes Phagocytic Ability of Embryonic Macrophages. Frontiers in Immunology, 9, 266.

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Immunolabeling of Neuronal Structures

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Brain slices were fixed as above, followed by 30% sucrose for cryoprotection. Thick sections (35 μm) were obtained with a sliding microtome (Microm, HM 400). Using standard immunocytochemical techniques, free-floating sections were rinsed twice in PBS buffer followed by 50% alcohol. Non-specific binding was blocked as above and sections were exposed to Fluorescein Avidin D (Vector Laboratories) for visualizing biocytin-filled cells followed by primary antisera at 4°C for 12–48 hours with vesicular GABA transporter (VGAT) rabbit polyclonal, a gift from Dr. R Reimer, and gephyrin mouse monoclonal (Synaptic Systems). Some sections were exposed to secondary antibody-conjugates with indocarbocyanine and/or indodicarbocyanine (Cy3 and Cy5; 1:200 Jackson ImmunoResearch Lab), Alexa Fluor 568 and/or Alexa Fluor 488 (both Alexa fluorophores, 1:1000; Invitrogen Corporation) for 2 hours at room temperature to allow triple labeling when necessary. Table 1 provides a list of antibodies used, sources and concentrations. The specificity of BDNF and TrkB receptor antibodies used in this study has been supported in previous studies (Snapyan et al., 2009 (link); Bergami et al., 2013 (link)).

Gu F., Parada I., Shen F., Li J., Bacci A., Graber K., Taghavi R.M., Scalise K., Schwartzkroin P., Wenzel J, & Prince D.A. (2017). Structural Alterations in Fast-Spiking GABAergic Interneurons in a Model of Posttraumatic Neocortical Epileptogenesis. Neurobiology of disease, 108, 100-114.

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Immunofluorescence Staining Protocol for α-Synuclein

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Immunofluorescence stainings were performed with sheep anti-TH (Abcam, cat# ab118 (working dilution 1:2000)), mouse anti α-syn (BD bioscience, cat# 610787 (working dilution 1:1000)) and rabbit anti-p129 α-syn (Abcam, Cat#51253 (working dilution 1:1000))
antibodies. The free-floating sections were washed thrice with PBS from anti-freeze solution.
The sections were then incubated in 0.025% Triton X-100 (Sigma Aldrich) in PBS containing 5% of normal serums (Vector Laboratories Inc, US) matching the species used to raise the secondary antibody for that proper stainings for 1 h. Primary antibodies were diluted in PBS and overnight incubated at 4 °C. The next day, the sections were washed thrice with PBS for 5 min, then incubated with Cy3 and Cy5 (working dilution 1:300) fluorophore conjugated secondary antibodies (Jackson Immunoresearch, Sweden). The sections were mounted on positive charged superfrost plus glass slides (ThermoScientific, US) and cover-slipped using PVA/DABCO. The immunofluorescence stainings were visualized on a Leica SP8 laserscanning confocal microscope (Leica, Germany).

Arkan S., Ljungberg M., Kirik D., & Hansen C. (2020). DNAJB6 Suppresses Alpha-synuclein Induced Pathology in an Animal Model of Parkinson’s Disease.

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