Cyanine 5

Manufactured by Jackson ImmunoResearch

Sourced in United States

Cyanine 5 is a fluorescent dye commonly used in molecular biology and biotechnology applications. It is a member of the cyanine dye family and has a maximum excitation wavelength of approximately 650 nm and a maximum emission wavelength of approximately 670 nm. Cyanine 5 can be used for labeling and detection of various biomolecules, such as proteins, nucleic acids, and other cellular components.

Automatically generated - may contain errors

Lab products found in correlation

Cyanine 3, by Jackson ImmunoResearch (3 mentions) Lsm 710, by Zeiss (2 mentions) Axiovert 200m, by Zeiss (2 mentions) Sirius red solution, by Merck Group (1 mentions) Ab1218, by Abcam (1 mentions) Anti vimentin, by Santa Cruz Biotechnology (1 mentions) Ab5320, by Abcam (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Alexa fluor 546 phalloidin, by Thermo Fisher Scientific (1 mentions) Zen black software, by Zeiss (1 mentions) Horse serum, by Thermo Fisher Scientific (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Phospho histone h3, by Abcam (1 mentions) Hoechst bisbenzimide h 33258, by Merck Group (1 mentions) Vt1000s vibrosector, by Leica (1 mentions) Superfrost slide, by Thermo Fisher Scientific (1 mentions) Neurod2, by Abcam (1 mentions) Proliferating cell nuclear antigen (pcna), by Merck Group (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Glycergel mounting medium, by Agilent Technologies (1 mentions)

Spelling variants of this product

Cyanine 5-conjugated streptavidin (2 citations)

5 protocols using cyanine 5

Muscle Tissue Immunostaining and Histological Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Muscle samples were fixed in 3.5% formaldehyde, embedded in paraffin and cut into 5 μm-thick sections. Sections were stained with an antibody against TWEAK (sc-5558, 1:100, SantaCruz Biotechnology). Bound primary antibodies were detected by immunofluorescence using secondary antibodies conjugated with Cyanine5 (Jackson ImmunoResearch Laboratories). Nuclei were stained with Bisbenzimid (Hoechst 33258). Control sections were treated similarly, except for the primary antibody, which was omitted.
For Ladewig staining, sections were brought to water with xylene and ethanol and treated with Ladewig's and Weigert's solution. For Sirius-red staining, sections were treated with Sirius-red solution (#365548, Sigma-Aldrich) for one hour and washed with acetic acid.
Digital evaluation and quantification were done on ImageJ software (National Institutes of Health, Bethesda, MD, USA.)

Al-Sawaf O., Fragoulis A., Rosen C., Kan Y.W., Sönmez T.T., Pufe T, & Wruck C.J. (2014). Nrf2 Protects Against TWEAK-mediated Skeletal Muscle Wasting. Scientific Reports, 4, 3625.

+ Open protocol

+ Expand

Antibody Validation and Characterization Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies purchased from commercial suppliers are as follows: FilaminA mouse monoclonal (AbNova, H00002316-M01, used at 1:1000); FilaminA mouse monoclonal (USBiological, F4510, used at 1:1000); NG2 mouse monoclonal (Upstate, 05-710, clone 132.38, used at 1:500); NG2 rabbit polyclonal (Chemicon, ab5320, used at 1:500); GFP mouse monoclonal (Abcam, ab-1218, used at 1:500); Rat anti-Myc tag antibody (JAC6-Serotec, used at 1:500). The mouse monoclonal antibodies agains α-tubulin and β-tubilin were a gift from Dr. Laurence Lafanachère, Institute for Advanced Biosciences, Grenoble, France (used at 1:5000 on western blot and at 1:1000 for IF). Mouse anti-Myc tag, A2B5, O4 hybridomas were amplified in the laboratory. Affinity-purified chicken and guinea pig antibody against RefilinA and RefilinB were obtained as previously described (Gay et al., 2011b (link)). Anti-Vimentin (Santa Cruz, sc-373717) was used at 1:1000. Alexa Fluor 546 Phalloidin (Invitrogen; A22283) was used at 1:250. Secondary antibodies: conjugated with Alexa Fluor 488 (Molecular Probes), Cyanine3 or Cyanine5 (Jackson ImmunoResearch) were all used at 1:2000. Secondary antibodies coupled to horseradish peroxidase were from Jackson ImmunoResearch (115-035-174, 111-035-144 and 112-035-175) and used at 1:5000.

Gay O., Gilquin B., Assard N., Stuelsatz P., Delphin C., Lachuer J., Gidrol X, & Baudier J. (2016). Refilins are short-lived Actin-bundling proteins that regulate lamellipodium protrusion dynamics. Biology Open, 5(10), 1351-1361.

+ Open protocol

+ Expand

Immunofluorescence Staining of Brain Sections

Check if the same lab product or an alternative is used in the 5 most similar protocols

Embryos were fixed by transcardiac perfusion with freshly-prepared 4% paraformaldehyde (Sigma). Brains were dissected, and 100 mm sections were prepared using a Leica VT1000S vibrosector. Slices were transferred into PBS with 0.5 μg/mL sodium azide (Sigma), then blocked with PBS supplemented with 3% horse serum (Invitrogen) and 0.3% Triton X-100 (Sigma) during 1 h, and incubated overnight at 4°C with the following primary antibodies: chicken GFP (Abcam, 1:2,000), rabbit Pax6 (Covance, 1:1000), Sox2 (Santa Cruz Biotechnology, 1:500), PCNA (Millipore, 1:200), phospho-Histone H3 (Abcam, 1:100), Tbr2 (Abcam, 1:500), Neurod2 (Abcam, 1:500), or mouse β3-tubulin (Tuj1 epitope, Covance, 1:1000). After three washes with PBS/0.1% Triton X-100, slices were incubated in PBS for 1 h at room temperature and incubated 2 h at room temperature with the appropriated Alexa 488 (1:1,000, Molecular Probes), Cyanine 3 or Cyanine 5 (1:400, Jackson ImmunoResearch) secondary antibodies. Sections were again washed three times with PBS/0.1% Triton X-100, stained with Hoechst (bisBenzimide H 33258, Sigma) for 5 min and washed twice in PBS. The sections were next mounted on a Superfrost slide (Thermo Scientific) and dried using a brush before adding Glycergel mounting medium (Dako). Imaging was performed using a Zeiss LSM780 confocal microscope controlled by the Zen Black software (Zeiss).

Bonnefont J., Tiberi L., van den Ameele J., Potier D., Gaber Z.B., Lin X., Bilheu A., Herpoel A., Velez Bravo F.D., Guillemot F., Aerts S, & Vanderhaeghen P. (2019). Cortical Neurogenesis Requires Bcl6-Mediated Transcriptional Repression of Multiple Self-Renewal-Promoting Extrinsic Pathways. Neuron, 103(6), 1096-1108.e4.

+ Open protocol

+ Expand

RNAscope-Based Multiplex Fluorescent Imaging

Check if the same lab product or an alternative is used in the 5 most similar protocols

The RNAscope TM Multiplex Fluorescent V2 kit (Biotechne, MN, USA) was used according to the manufacturer's instructions. All laboratory solutions were prepared in advance with autoclaved water at 0.1% DEPC (Sigma-Aldrich, MO, USA; D5758) to avoid any contamination by RNAses and DNAses. Our target was revealed with the Plat probe, revealing the plat gene encoding tPA (Biotechne, MN, USA; 586951) coupled with Opal 520 fluorophore (Akoya, MA, USA; SKU FP1487001KT; 1/5000). At the end of the protocol, we carried out immunostaining steps with the primary antibodies incubated overnight at 4 °C (anti-CD31 antibody, BD Biosciences; 555024 1/500; anti-phalloidin antibody, Invitrogen N21479 1/300). After 3 washes in 1 × PBS, the appropriate secondary antibodies coupled with Cyanine 3 or Cyanine 5 (1/800 Jackson Immunoresearch, West Grove, CA, USA) were incubated at room temperature for 90 min. Once washed, the slides were mounted with coverslip and mounting medium with DAPI (Fluoromont-G; Thermofisher, MA USA; 15596276).

Furon J., Yetim M., Pouettre E., Martinez de Lizarrondo S., Maubert E., Hommet Y., Lebouvier L., Zheng Z., Ali C, & Vivien D. (2023). Blood tissue Plasminogen Activator (tPA) of liver origin contributes to neurovascular coupling involving brain endothelial N-Methyl-D-Aspartate (NMDA) receptors. Fluids and Barriers of the CNS, 20, 11.

+ Open protocol

+ Expand

Fluorescent Labeling and Imaging of Exosome Uptake in Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

N2a stably overexpressing GFP-CD63 were fixed at room temperature for 20 min using 4% paraformaldehyde (PFA) in PBS, permeabilized 30 min with 0.05% saponin in PBS-3% BSA (Bovine Serum Albumin, Sigma). Labelling with mouse anti-LBPA antibody was revealed using Cyanin 5-labelled anti-mouse antibody (Jackson Immunoresearch 0.2 µg/ml). Primary and secondary antibodies were diluted in PBS-3% BSA.
After incubation with exosomes, neurons were fixed using 4% PFA, 4% sucrose in PBS. After extensive washes in PBS, cells were permeabilized for 10 min at 4°C using PBS containing 0.1% Triton X-100. Cells were incubated in PBS containing 3% goat pre-immune serum (GPI, Sigma-Aldrich) and stained with the indicated primary antibodies and with secondary antibodies conjugated to either Alexa Fluor 488, Alexa Fluor 594 or Cyanine 5 (Jackson Immunoresearch 0.2 µg/ml). Primary and secondary antibodies were diluted in PBS 1% GPI. Nuclei were stained with Hoechst (Sigma).
Coverslips were mounted in Mowiol (Calbiochem) and observed using epifluorescence (Axiovert 200 M, Zeiss) or confocal microscopy (LAS AF – TCS SPE, Leica or LSM 710, Zeiss using Zen software). Some figures show acquisition in one given plan, others present maximal intensity projections (cf. figure legends). Image files were processed with ImageJ.

Chivet M., Javalet C., Laulagnier K., Blot B., Hemming F.J, & Sadoul R. (2014). Exosomes secreted by cortical neurons upon glutamatergic synapse activation specifically interact with neurons. Journal of Extracellular Vesicles, 3, 10.3402/jev.v3.24722.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!