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Brdu assay kit

Manufactured by Cell Signaling Technology
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The BrdU Assay Kit is a laboratory tool used to detect and quantify cell proliferation. It measures the incorporation of the thymidine analog bromodeoxyuridine (BrdU) into the DNA of dividing cells. The kit provides the necessary reagents to perform this assay.

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Lab products found in correlation

Anti hdac1, by Cell Signaling Technology (1 mentions) Phytohemagglutinin (pha), by Merck Group (1 mentions) Anti phospho iκbα, by Cell Signaling Technology (1 mentions) Anti nf κb p65, by Cell Signaling Technology (1 mentions) Anti iκbα, by Cell Signaling Technology (1 mentions) Anti gapdh, by Abcam (1 mentions) Eon microplate spectrophotometer, by Agilent Technologies (1 mentions) Synergy 2 multi detection microplate reader, by Agilent Technologies (1 mentions) Glomax discover multiplate reader, by Promega (1 mentions)

Close spelling variants of this product

ELISA (BrdU Cell Proliferation Assay Kit, 5-bromo-2′-deoxyuridine (BrdU) cell proliferation assay kit 5-Bromo-2′-deoxyuridine (BrdU) incorporation assay kit BrdU Cell Proliferation Assay Kit BrdU) proliferation assay kit BrdU incorporation assay kit BrdU ELISA assay kit BrdU Cell Proliferation Chemiluminescent Assay Kit BrdU assay

9 protocols using brdu assay kit

1

Dasatinib Modulates BV2 Microglial Proliferation

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To investigate the effects of dasatinib on the proliferation of BV2 microglial cells via a non-metabolic assay, a BrdU assay kit (Cell Signaling, Danvers, MA, USA) was used. BV2 microglial cells were seeded in 96-well plates at a density of 4 × 104 cells/well and treated with various concentrations of dasatinib (100, 250, 500, 750, and 1000 nM) for 24 h in the absence of FBS. In addition, BV2 microglial cells were seeded in 96-well plates and treated with dasatinib (250 nM) or vehicle (1% DMSO) for 30 min followed by LPS (200 ng/ml) or PBS for 23.5 h. The BrdU assays were conducted in accordance with the manufacturer’s instructions. The labeling time with BrdU (10 μM) was 4 h, and BrdU incorporation was detected by an anti-BrdU antibody and HRP-conjugated secondary antibody. The absorbance was measured at 450 nm. In addition, we conducted parallel experiments with mouse primary astrocytes in the presence/absence of LPS treatment using the BrdU assay.
Ryu K.Y., Lee H.J., Woo H., Kang R.J., Han K.M., Park H., Lee S.M., Lee J.Y., Jeong Y.J., Nam H.W., Nam Y, & Hoe H.S. (2019). Dasatinib regulates LPS-induced microglial and astrocytic neuroinflammatory responses by inhibiting AKT/STAT3 signaling. Journal of Neuroinflammation, 16, 190.
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2

Investigating 17-OHPC and Immune Modulation

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17-OHPC was synthesized by Symbiotec Pharmalab (Lot # ZHPCy19003) and provided by Evergreen Therapeutics, Inc. for the studies, castor oil (Sigma-Aldrich, #C9606) and 50% ethanol in PBS were used as the vehicles for in vivo and in vitro experiments, respectively. Muromonab-CD3 (OKT3) and anti-CD28 antibody were from Takara (#T210) and Sigma-Aldrich (# 217669), respectively. Phytohemagglutinin (PHA) was obtained from Sigma-Aldrich (# 11249738001). BrdU assay kit was from Cell Signaling Technology (# 6813). The antibodies used in this study included anti-phospho-IκBα (# 2859), anti-IκBα (# 9242), anti-NF-κB p65 (# 8242), anti-HDAC1 (# 5356) from Cell Signaling Technology, and anti-GAPDH (# ab9485) from Abcam. Unless otherwise specified, all cell culture reagents were purchased from Sigma-Aldrich or Life Technologies.
Hu T., Tang C., Stern S., Yang L, & Du T. (2022). 17α-Hydroxyprogesterone Caproate Inhibits Cytokine Production via Suppression of NF-κB Activation. Frontiers in Pharmacology, 13, 831315.
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3

Glucose and Fructose Effects on Cell Proliferation

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IMKC were plated in 96 well plates with a seeding density of 2.5 × 104 cells/well in culturing RPMI. Cells were then treated with 25 mM glucose or fructose for 24 h. BrdU reagent was added at time of treatment using the BrdU Assay Kit (#6813 Cell signaling technology) and absorbance measured at 450 nm on plate reader.
Lodge M., Scheidemantle G., Adams V.R., Cottam M.A., Richard D., Breuer D., Thompson P., Shrestha K., Liu X, & Kennedy A. (2024). Fructose regulates the pentose phosphate pathway and induces an inflammatory and resolution phenotype in Kupffer cells. Scientific Reports, 14, 4020.
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4

BrdU Assay for HDF DNA Synthesis

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To quantify the DNA synthesis of HDFs under the influence of HSA-bFGF NPs, BrdU assay kit (Cell Signaling Technology, USA) was used. When cells were cultured with BrdU-containing media, the pyrimidine, a type of nucleotide analog, incorporated into DNA replacing thymidine. After HDFs were seeded onto 48-well plate, 20 µg/ml of control HSA NPs, 5 µg/ml of commercial bFGF, 5 µg/ml of bFGF, and 20 µg/ml of HSA-bFGF NPs were treated to the cells, respectively. The BrdU assay was performed at day 2 and 4. After adding 10 µM of BrdU solution to cell culture medium, HDFs were incubated at 37 ℃ for 3 h in order to induce incorporation of BrdU into cellular DNA. Then, the subsequent procedures were performed according to the manufacturer’s instructions. The absorbance was measured using microplate reader (BioTek™ Eon™ Microplate Spectrophotometers) at 450 nm.
Son B., Kim M., Won H., Jung A., Kim J., Koo Y., Lee N.K., Baek S.H., Han U., Park C.G., Shin H., Gweon B., Joo J, & Park H.H. (2023). Secured delivery of basic fibroblast growth factor using human serum albumin-based protein nanoparticles for enhanced wound healing and regeneration. Journal of Nanobiotechnology, 21, 310.
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5

Quiescent and Cycling Cell BrdU Assay

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EGFP-p27highmCherry-Ki67low quiescent (Q) and cycling (C) cells seeded in 96-well plates (5 × 103 cells per well) were subjected to DNA synthesis assays using the BrdU Assay kit (Cell Signalling) as described previously 22 (link). Absorbance was read at 450 nm using a Synergy™ 2 multidetection microplate reader (BioTek, VT).
La T., Chen S., Guo T., Zhao X.H., Teng L., Li D., Carnell M., Zhang Y.Y., Feng Y.C., Cole N., Brown A.C., Zhang D., Dong Q., Wang J.Y., Cao H., Liu T., Thorne R.F., Shao F.M., Zhang X.D, & Jin L. (2021). Visualization of endogenous p27 and Ki67 reveals the importance of a c-Myc-driven metabolic switch in promoting survival of quiescent cancer cells. Theranostics, 11(19), 9605-9622.
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6

Cell Proliferation Assays Protocols

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The following kits and reagents were used in our experiments. The X-Gal assay kit was obtained from Promega, the BrdU assay kit was obtained from Cell Signaling Technology, and the Click EdU imaging kit was obtained from Thermo Fisher. All kits were used according to the manufacturer’s instructions. BrdU absorbance was measured with a GloMax Discover multiplate reader (Promega).
Middonti E., Astanina E., Vallariello E., Hoza R.M., Metovic J., Spadi R., Cristiano C., Papotti M., Allavena P., Novelli F., Parab S., Cappello P., Scarpa A., Lawlor R., Di Maio M., Arese M, & Bussolino F. (2024). A neuroligin-2-YAP axis regulates progression of pancreatic intraepithelial neoplasia. EMBO Reports, 25(4), 1886-1908.
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7

BrdU Cell Proliferation Assay

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Cell proliferation was concurrently analyzed using a BrdU assay kit (Cell Signaling Technology, Inc., Danvers, MA, USA). The steps were performed according to the manufacturer's protocol. In brief, HGC-27 cells were incubated at 37°C with 20 µM BrdU for 40 min prior to being harvested. Subsequently, the cells were treated with 1× fixing solution (cat no. 6813; Cell Signaling Technology, Inc.) at room temperature for 30 min, followed by incubation with anti-BrdU antibody (cat no. 6813; 1:1,000; Cell Signaling Technology, Inc.) at 37°C for 1 h. Finally, the optical absorbance was measured with a multimode microplate reader at dual wavelengths of 450/550 nm.
Li Y., Gao W., Ma Y., Zhu G., Chen F, & Qu H. (2018). Dual targeting of survivin and X-linked inhibitor of apoptosis protein suppresses the growth and promotes the apoptosis of gastric cancer HGC-27 cells. Oncology Letters, 16(3), 3489-3498.
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8

Podocyte Proliferation Assay Protocol

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Proliferation rate was assessed using a colorimetric BrdU assay kit (#6813, Cell signaling) according to the manufacturer’s instructions. Experiments were performed in human immortalized podocytes 7 days after retroviral shRNA knockdown of OSGEP, TP53RK, or TRPKB (passage 4). BrdU incorporation is measured as absorbance at 450 nm using a spectrophotometer. Data is plotted as mean and standard deviation. Statistical significance was calculated using one-way ANOVA multi-test analysis with post hoc testing according to Sidak. P values <0.01 and <0.05 were considered statistically significant and are indicated in the figures. Measurements were performed in triplicates, and experiments were repeated twice independently.
Braun D.A., Rao J., Mollet G., Schapiro D., Daugeron M.C., Tan W., Gribouval O., Boyer O., Revy P., Jobst-Schwan T., Schmidt J.M., Lawson J.A., Schanze D., Ashraf S., Boddaert N., Collinet B., Martin G., Liger D., Lovric S., Furlano M., Guerrera I.C., Sanchez-Ferras O., Menten B., Vergult S., De Rocker N., Airik M., Hermle T., Shril S., Widmeier E., Gee H.Y., Choi W.I., Sadowski C.E., Pabst W.L., Warejko J., Daga A., LeBerre T.B., Matejas V., Behnam B., Beeson B., Begtrup A., Bruce M., Ch'ng G.S., Lin S.P., Chang J.H., Chen C.H., Cho M.T., Gipson P.E., Hsu C.H., Kari J.A., Ke Y.Y., Kiraly-Borri C., Lai W.M., Lemyre E., Littlejohn R.O., Masri A., Moghtaderi M., Nakamura K., Praet M., Prasad C., Prytula A., Roeder E., Rump P., Schnur R.E., Shiihara T., Sinha M., Soliman N.A., Soulami K., Sweetser D.A., Tsai W.H., Tsai J.D., Vester U., Viskochil D.H., Vatanavicharn N., Waxler J.L., Wolf M.T., Wong S.N., Poduri A., Truglio G., Mane S., Lifton R.P., Bouchard M., Kannu P., Chitayat D., Magen D., Calleweart B., van Tilbeurgh H., Zenker M., Antignac C, & Hildebrandt F. (2017). Mutations in the evolutionarily highly conserved KEOPS complex genes cause nephrotic syndrome with microcephaly. Nature genetics, 49(10), 1529-1538.
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9

Cell Proliferation Assay for LAC117

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To measure the cell proliferation activity of LAC117 in Huh7 and HepG2 cells, 8 × 103 cells were plated per well onto 96-well plates. Following overnight culture, LAC117 was added at specified concentrations. After 24 h of incubation, cell proliferation was measured with a BrdU assay kit (Cell Signaling, cat.n.6813) per the manufacturer’s instructions. Plates were read at 450 nm by using a spectrometer.
Kim J., Jung K.H., Yan H.H., Cheon M.J., Kang S., Jin X., Park S., Oh M.S, & Hong S.S. (2018). Artemisia Capillaris leaves inhibit cell proliferation and induce apoptosis in hepatocellular carcinoma. BMC Complementary and Alternative Medicine, 18, 147.
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