Cy3 conjugated antibodies

Manufactured by Jackson ImmunoResearch

Sourced in United States

Cy3-conjugated antibodies are fluorescently-labeled secondary antibodies used for immunodetection applications. They are comprised of a Cy3 dye molecule covalently attached to the antibody. The Cy3 fluorophore has excitation and emission wavelengths optimized for visualization and quantification purposes.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Nis element, by Nikon (1 mentions) Ago2 antibody, by Abcam (1 mentions) Dm6000, by Leica (1 mentions) Mouse anti α catenin, by BD (1 mentions) Rabbit anti β catenin, by Merck Group (1 mentions) Anti gfp, by Thermo Fisher Scientific (1 mentions) Ketamine, by Zoetis (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Anti tdp 43, by Abnova (1 mentions) Anti c myc, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Cy3- and Cy5-conjugated secondary antibodies (10 citations) Cy2 and Cy3 conjugated secondary antibodies (10 citations) Cy3-conjugated IgG antibodies (5 citations) Cy3- or Alexa 488-conjugated secondary antibodies (5 citations) Cy3-conjugated and Alexa 488-conjugated secondary antibodies (3 citations) Cyanine 3 (Cy3)-conjugated secondary antibodies (2 citations) Cy3-conjugated goat-α-rabbit antibodies (2 citations) Secondary antibodies conjugated with Cy3 (2 citations) Cy3-conjugated anti-rabbit or anti-mouse secondary antibodies (2 citations) Cyanine 3 (Cy3)- and FITC-conjugated secondary antibodies (2 citations)

4 protocols using cy3 conjugated antibodies

Immunofluorescence Analysis of EAAT3 in IPEC-J2 Cells

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pcDNA3.1+ and EAAT3-pcDNA3.1+ stably transfected IPEC-J2 cells were seeded at 1×10⁴ cells/mL into 96-well culture plates and fixed with 4% paraformaldehyde for 30 min. Cells were then permeabilized with 0.3% Triton X-100 for 15 min and blocked in a protein solution (Dako, Carpinteria, CA) for 20 min. The primary antibody, anti-EAAT3 (1:500 in antibody diluent; Dako, Carpinteria, CA), was applied to these cells for 2 h at room temperature. Secondary staining was performed with Cy3 conjugated antibodies (Jackson ImmunoResearch, West Grove, PA) (1:200 in antibody diluent) with incubation at room temperature for 1 h. Nuclei were stained with 4, 6 diamidino-2-phenylindole (DAPI, 1:1000 in PBS; Sigma, St. Louis, USA), for 5 min at room temperature. Fluorescence signals were observed with a fluorescence microscope (NIS-Elements, Nikon, Japan). At least three independent experiments were performed to verify results.

Ye J.L., Gao C.Q., Li X.G., Jin C.L., Wang D., Shu G., Wang W.C., Kong X.F., Yao K., Yan H.C, & Wang X.Q. (2016). EAAT3 promotes amino acid transport and proliferation of porcine intestinal epithelial cells. Oncotarget, 7(25), 38681-38692.

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Immunostaining of AGO2 and p62 in Cells

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Following treatments with various stimuli, cells were washed with PBS, fixed for 20 minutes in 4% paraformaldehyde in PBS, permeabilized by incubation in 0.1% Triton X-100 in PBS for 20 minutes, and blocked for 1 hour with 5% milk in PBS. Permeabilized cells were incubated overnight at 4°C with primary antibody diluted in 5% milk in PBS: 1:200 rabbit polyclonal AGO2 antibody (Abcam, Toronto, Canada), and 1:1000 rabbit polyclonal p62 antibody (Santa Cruz Biotechnology, Santa Cruz, CA). Cells were then washed with PBS, and incubated for 1 hour at room temperature with secondary Cy3-conjugated antibodies (Jackson ImmunoResearch, West Grove, PA). Confocal image acquisition was performed in a Quorum Spinning Disk Confocal Microscope.

Sibony M., Abdullah M., Greenfield L., Raju D., Wu T., Rodrigues D.M., Galindo-Mata E., Mascarenhas H., Philpott D.J., Silverberg M.S, & Jones N.L. (2015). Microbial Disruption of Autophagy Alters Expression of the RISC Component AGO2, a Critical Regulator of the miRNA Silencing Pathway. Inflammatory Bowel Diseases, 21(12), 2778-2786.

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Immunofluorescent Staining of Catenins

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Cells were fixed for 12 min at room temperature using 4% paraformaldehyde in PBS, and then rinsed with PBS and permeabilized for 45 min in PBS supplemented with 1.5% BSA and 0.1% Triton X-100. Cells were then incubated for 1 h with mouse anti–α-catenin (BD) at 1:400 or rabbit anti–β-catenin (Sigma-Aldrich) at 1:400 dilution in PBS-BSA, rinsed, and incubated 1 h with anti–mouse or anti–rabbit Cy3–conjugated antibodies (Jackson ImmunoResearch Laboratories, Inc.) at 1:500 dilution. Preparations were mounted in Mowiol, 90% glycerol, and PBS. Images were taken with a microscope (DM6000; Leica) equipped with a 63× oil objective and Micromax charge coupled device (CCD) camera (Roper Scientific).

Strale P.O., Duchesne L., Peyret G., Montel L., Nguyen T., Png E., Tampé R., Troyanovsky S., Hénon S., Ladoux B, & Mège R.M. (2015). The formation of ordered nanoclusters controls cadherin anchoring to actin and cell–cell contact fluidity. The Journal of Cell Biology, 210(2), 333-346.

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Immunostaining Neurodegeneration Markers

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Animals were anesthetized using a cocktail of xylazine (20 mg ml⁻¹; Butler, Columbus, OH, USA), ketamine (100 mg ml⁻¹; Fort Dodge Animal Health, Fort Dodge, IA, USA) and acepromazine (10 mg ml⁻¹; Boerhinger Ingelheim, St Joseph, MO, USA) in a 3:3:1 fluid ratio. Animals were administered the cocktail intramuscularly at a dose of 1 ml kg⁻¹ and perfused with phosphate-buffered saline followed by cold 4% paraformaldehyde in phosphate-buffered saline. Tissues were removed and immersed in 4% paraformaldehyde overnight at 4 °C. Fifty-micrometer sections were cut on a sliding microtome with a freezing stage. Primary antibodies used were anti-TDP-43 (Abnova), anti-c-myc (Santa Cruz Biotechnology) and anti-GFP (Invitrogen) at dilutions of 1:500 and 1:1000. Secondary antibodies include biotinylated antibodies from DAKO Cytomation (Carpinteria, CA, USA; 1:2000) and Alexa Fluor 488-(Invitrogen) or Cy3-conjugated antibodies (Jackson ImmunoResearch, West Grove, PA, USA) at 1:300. 4′,6-Diamidino-2-phenylindole (Sigma, St Louis, MO, USA) counterstaining and Nissl staining followed standard methods.

Jackson K., Dayton R., Orchard E., Ju S., Ringe D., Petsko G., Maquat L, & Klein R. (2014). Preservation of forelimb function by UPF1 gene therapy in a rat model of TDP-43-induced motor paralysis. Gene therapy, 22(1), 20-28.

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