Cell proliferation dye 450

Manufactured by Thermo Fisher Scientific

Cell proliferation dye 450 is a fluorescent dye used to track and measure cell division in vitro. It binds to cellular proteins and is partitioned equally between daughter cells during cell division, allowing quantification of proliferation.

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Lab products found in correlation

Facscanto 2 flow cytometer, by BD (1 mentions) Efluor 450, by Thermo Fisher Scientific (1 mentions) Fixation permeabilization kit, by BD (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Imdm media, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Cell Proliferation Dye eFluor 450 (69 citations) Cell proliferation dye (10 citations) EBioscience™ cell proliferation dye eFluor 450 (10 citations) EFluor 450 cell proliferation dye (CPD, (5 citations)

2 protocols using cell proliferation dye 450

Multiparameter Flow Cytometry Analysis

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The antibodies used for FACS analysis are summarized in Table 1. The efluor 450-labeled anti-Foxp3 staining was performed using the eBioscience kit and protocol. The fixation/permeabilization kit (BD Biosciences) was used for intra-cellular staining of active caspase-3. Cell proliferation dye 450 (ebioscience) was used to help exclude APCs in the in vivo experiment. Flow cytometry events were acquired on a FACSCanto II flow cytometer (BD Biosciences) and analyzed using FlowJo (Tree Star, Ashland, OR, USA) software.

Pilon C., Stehlé T., Beldi-Ferchiou A., Matignon M., Thiolat A., Burlion A., Grondin C., Birebent B., Pirenne F., Rouard H., Lang P., Marodon G., Grimbert P, & Cohen J.L. (2019). Human Apoptotic Cells, Generated by Extracorporeal Photopheresis, Modulate Allogeneic Immune Response. Frontiers in Immunology, 10, 2908.

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CD4+ T Cell Proliferation Analysis

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Upon isolation, 2.5×10⁶ primary CD4⁺ T cells were labeled with cell proliferation dye 450 (eBiosciences). Briefly cells were washed two times with PBS (Life Technologies) and stained in 1× dye in a 2.0 × 10⁷ concentration for 20 minutes at room temperature in the dark. The reaction was quenched by adding 5× IMDM media (Life Technologies), and incubated on ice for 5 min in the dark. Cells were washed twice with media, and plated at a concentration of 0.6×10⁶ cells and cultured as described above for activation and polarization. On day 3, cells were split and cultured as described above. On day 5, 1×10⁶ cells were additionally stained with CD4 (GK1.5, Biolgend) and analyzed by flow cytometry.

Chisolm D.A., Savic D., Moore A.J., Ballesteros-Tato A., León B., Crossman D.K., Murre C., Myers R.M, & Weinmann A.S. (2017). CCCTC-Binding Factor Translates Interleukin 2- and α-Ketoglutarate-Sensitive Metabolic Changes in T Cells into Context-Dependent Gene Programs. Immunity, 47(2), 251-267.e7.

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