Brdu flow kit protocol

Manufactured by BD

The BrdU flow kit is a laboratory reagent used to detect and measure cell proliferation. It provides a fluorescent-based method for detecting bromodeoxyuridine (BrdU) incorporation into newly synthesized DNA, which is an indicator of cellular division. The kit contains the necessary reagents and protocols to perform this analysis using flow cytometry.

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Lab products found in correlation

Anti brdu, by BD (2 mentions) Anti cd4, by Thermo Fisher Scientific (1 mentions) Anti cd11c, by Thermo Fisher Scientific (1 mentions) Anti cd8α, by Thermo Fisher Scientific (1 mentions) Anti f4 80 bm8, by BioLegend (1 mentions) Anti cd5 53 7.3, by BD (1 mentions) Anti cd19 ebio 1d3, by Thermo Fisher Scientific (1 mentions) Lsrii cytometer, by BD (1 mentions) Bromodeoxyuridine (brdu), by BD (1 mentions) Fxcycle pi rnase staining solution, by BD (1 mentions)

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BrdU Flow Kit (453 citations)

3 protocols using brdu flow kit protocol

BrdU Uptake in Immune Cells

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BrdU was delivered to mice through drinking water at the concentration of 1mg/ml + 1% glucose. Spleen and lymph nodes were harvested on day 6 post-immunization or infection. Following tetramer enrichment, cells were stained with cell surface antibodies [anti-CD3, anti-CD4, anti-CD19 (eBio (1D3), eBiosciences), anti-CD8α, anti-CD11c (eBiosciences), anti-F4/80 (BM8, Biolegend) and anti-CD5 (53–7.3, BD Pharmingen)] followed by intracellular anti-BrdU (BD Pharmingen) antibody according to the BrdU flow kit protocol (BD Biosciences).

Deshpande N.R., Uhrlaub J.L., Way S.S., Nikolich-Žugich J, & Kuhns M.S. (2018). A disconnect between precursor frequency, expansion potential, and site-specific CD4+ T cell responses in aged mice. PLoS ONE, 13(6), e0198354.

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BrdU Labeling of Immune Cells

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BrdU was administered to mice through drinking water at the concentration of 1 mg/ml + 1% glucose. Spleen and lymph nodes were harvested on day 6. Post-tetramer enrichment, cells were stained with cell surface antibodies (anti-CD3, anti-CD4, anti-CD19, anti-CD8α, anti-CD11c, anti-F4/80, anti-CD44, and anti-CD5) followed by intracellular anti-BrdU (BD Pharmingen) antibody according to BrdU flow kit protocol (BD Biosciences).

Deshpande N.R., Parrish H.L, & Kuhns M.S. (2015). Self-recognition drives the preferential accumulation of promiscuous CD4+ T-cells in aged mice. eLife, 4, e05949.

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BrdU Labeling and Cell Cycle Analysis

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KPR cells and KPR^p53React cells were plated on day 0 in complete RPMI-1640 medium supplemented with 10% fetal bovine serum. After 24 h cells were pulsed with BrdU according to manufacturer instructions (Becton Dickinson). BrdU was removed and replaced with fresh medium. On day 2, cells were exposed to tamoxifen to restore p53 (KPR^p53). On day 4, cells were stained with CellTrace Violet as described above while attached to the plate to minimize handling. On day 10, cells were harvested and stained with an anti-BrdU PE-conjugated antibody according to the manufacturer’s BrdU Flow Kit protocol (BD Biosciences). For cell cycle analysis, cells were fixed in 70% ethanol and stained with FxCycle PI/RNase staining solution (BD Biosciences) according to manufacturer’s instruction. Samples were analyzed using an BD^™ LSRII cytometer (Becton Dickinson) and data were analyzed by FlowJo 10.5 software (Becton Dickinson).

Perego M., Tyurin V.A., Tyurina Y.Y., Yellets J., Nacarelli T., Lin C., Nefedova Y., Kossenkov A., Liu Q., Sreedhar S., Pass H., Roth J., Vogl T., Feldser D., Zhang R., Kagan V.E, & Gabrilovich D.I. (2020). Reactivation of Dormant Tumor Cells by Modified Lipids Derived from Stress-Activated Neutrophils. Science translational medicine, 12(572), eabb5817.

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