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Brdu elisa assay

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The BrdU ELISA assay is a laboratory kit used to detect and quantify the incorporation of the synthetic thymidine analog bromodeoxyuridine (BrdU) into cellular DNA. This assay provides a measure of cell proliferation and can be used in a variety of cell-based applications.

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Lab products found in correlation

Evos microscope, by Thermo Fisher Scientific (2 mentions) Xtt assay, by Merck Group (1 mentions) Prism 6, by GraphPad (1 mentions) Glutamax 1, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Mitomycin c, by Merck Group (1 mentions) Bromodeoxyuridine (brdu), by Merck Group (1 mentions) Synergy ht microplate reader, by Agilent Technologies (1 mentions) Caspase glo 3 7 assay, by Promega (1 mentions) Crystal violet, by Merck Group (1 mentions) Dulbecco s modified eagle s medium high glucose, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) N hexane, by Merck Group (1 mentions) N butanol, by Merck Group (1 mentions) Infinite m200, by Tecan (1 mentions)

14 protocols using brdu elisa assay

1

Cell Viability and Proliferation Assay

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For the cell viability assay, 1 × 104 cells were seeded per well in a 96-well plate and incubated overnight and then treated with various concentrations of reagents. After 24 h, the growth medium was discarded, and MTT assay solution was added for 1 h at 37 °C. Newly formed mitochondrial MTT crystals were dissolved with dimethyl sulfoxide, and the absorbance was read using a microplate reader. For the cell proliferation assay, a BrdU ELISA assay (Roche Applied Science) was used to measure the rate of DNA synthesis.
Lee C.C., Lin J.C., Hwang W.L., Kuo Y.J., Chen H.K., Tai S.K., Lin C.C, & Yang M.H. (2018). Macrophage-secreted interleukin-35 regulates cancer cell plasticity to facilitate metastatic colonization. Nature Communications, 9, 3763.
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2

Cytotoxicity and Senescence Assay of DS-3032b

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Cells were seeded in triplicate onto 96-well plates, incubated for 24 h to permit adherence, then treated with 0 - 2000 nM DS-3032b for 24, 48 or 72 h. Cell viability was determined using the 2,3-Bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay (Sigma-Aldrich) according to the manufacturer's protocol. The IC50 was calculated using GraphPad Prism 6.0 (GraphPad Software). Cell proliferation was determined using the BrdU ELISA assay (Roche) according to the manufacturer's protocol. Senescence was measured using the fluorometric SA-β-gal activity assay (Cell Biolabs Inc.) according to the manufacturer's protocol and corrected for cell viability (XTT assay).
Arnhold V., Schmelz K., Proba J., Winkler A., Wünschel J., Toedling J., Deubzer H.E., Künkele A., Eggert A., Schulte J.H, & Hundsdoerfer P. (2017). Reactivating TP53 signaling by the novel MDM2 inhibitor DS-3032b as a therapeutic option for high-risk neuroblastoma. Oncotarget, 9(2), 2304-2319.
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3

Analyzing MSC Immunomodulatory Effects in MLR

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Standard 5-day MLR cultures were set up with 5 × 104 Mitomycin C–treated (Sigma-Aldrich) human peripheral blood mononuclear cells (PBMCs) as stimulators and 2 × 105 human CD4+ T-cells (Lonza) in 96-well round-bottom plates in 200 μL complete medium consisting RPMI 1640 (Life Technologies) supplemented with 0.1 mM β-mercaptoethanol, 10% FBS, GLUTAMAX I (Life Technologies), 100 U/mL penicillin, and 100 μg/mL streptomycin in the presence or absence of iPSC-MSCs and BM-MSCs. For analyzing expression of CD69+ and CD25+ regulatory T-cell population, 106 responder cells were mixed with 2.5 × 105 stimulator PMNCs in presence or absence of 2 × 105 iPSC-MSCs or BM-MSCs. MLRs were performed on a layer of confluent Mitomycine C treated MSCs seeded one day before. Proliferation was determined with BrdU ELISA assay (Roche) based on manufacturer instruction. IL-2 and IFN-γ concentration was determined in MSC/MLR coculture supernatants using a commercially available ELISA (BD Bioscience) according to manufacturer's instructions. CD25 and CD69 (BD Bioscience) expression on CD4+ cells were analyzed by flow cytometry.
Moslem M., Eberle I., Weber I., Henschler R, & Cantz T. (2015). Mesenchymal Stem/Stromal Cells Derived from Induced Pluripotent Stem Cells Support CD34pos Hematopoietic Stem Cell Propagation and Suppress Inflammatory Reaction. Stem Cells International, 2015, 843058.
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4

Chemokine-induced cell proliferation

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Cells were plated in 96-well plates, and treated with 250 ng/mL mCcl5 (R&D systems, 478-MR-025) or 0.5 % BSA in PBS (vehicle control) for 4 hours at 37°C. BrdU was added to the cell culture 2 hours before fixing the cells and performing the BrdU ELISA assay (Roche Life Science, 11647229001) according the manufacturer's protocol.
Pan Y., Smithson L.J., Ma Y., Hambardzumyan D, & Gutmann D.H. (2017). Ccl5 establishes an autocrine high-grade glioma growth regulatory circuit critical for mesenchymal glioblastoma survival. Oncotarget, 8(20), 32977-32989.
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5

Platelet-Induced Cell Proliferation

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Cell proliferation was assessed by BrdU incorporation using colorimetric BrdU ELISA assay (Roche, Rotkreuz, Switzerland) according to the manufacturer’s protocol. MCECs (2 × 103 per well) were stimulated by isolated platelets (2 × 107 per well).
Jank M., von Niessen N., Olivier C.B., Schmitt H., Anto-Michel N., Hilgendorf I., Bode C., Moser M., Esser J.S, & Zhou Q. (2021). Platelet Bone Morphogenetic Protein-4 Mediates Vascular Inflammation and Neointima Formation after Arterial Injury. Cells, 10(8), 2027.
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6

Cell Proliferation Evaluation of Phytotherapeutics

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To quantify the effects of EGCG, silibinin and noscapine on cell proliferation, a colorimetric cell proliferation 5-bromo-2′-deoxyuridine (BrdU)-ELISA assay (Roche Diagnostics GmbH, Mannheim, Germany) was performed according to the manufacturer's protocol. Briefly, cells were seeded at 1×104 cells/well in 96-well plates and cultured for 24 h. The phytotherapeutic agents were subsequently added in the appropriate concentrations for 24 h. The BrdU labelling solution was added and incubated for a further 24 h. BrdU, a pyrimidine analogue, integrates into the DNA of proliferating cells. The level of proliferation was quantified by the light emission detected via an Orion Microplate Luminometer (Berthold Detection Systems GmbH, Pforzheim, Germany). Cell proliferation was determined in quadruplicate. The results are expressed as a percentage of the proliferation of DMSO-treated control cells.
Harati K., Behr B., Wallner C., Daigeler A., Hirsch T., Jacobsen F., Renner M., Harati A., Lehnhardt M, & Becerikli M. (2016). Anti-proliferative activity of epigallocatechin-3-gallate and silibinin on soft tissue sarcoma cells. Molecular Medicine Reports, 15(1), 103-110.
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7

BrdU Proliferation and Caspase 3/7 Apoptosis Assay

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Cells were seeded in 96-well plates (20,000 cells/well): 24 h after seeding, cells were pretreated with BafA1 for 4 h, and after that, bromodeoxyuridine (BrdU, Sigma-Aldrich) was added to the culture medium for the next 24 h. Proliferation was measured as BrdU incorporation using the colorimetric cell proliferation BrdU ELISA assay (Roche) according to the manufacturer’s protocol. Apoptosis was analyzed by measuring caspase 3/7 activity using the Caspase-Glo 3/7 assay (Promega, Dubendort, Switzerland) following the manufacturer’s protocol. BrdU absorbances and Cas3/7 luminescence were measured with the Synergy HT microplate reader (BioTek, Zurich, Switzerland). Each sample was measured in quadruplicates.
Seidenberg J., Stellato M., Hukara A., Ludewig B., Klingel K., Distler O., Błyszczuk P, & Kania G. (2021). The AP-1 Transcription Factor Fosl-2 Regulates Autophagy in Cardiac Fibroblasts during Myocardial Fibrogenesis. International Journal of Molecular Sciences, 22(4), 1861.
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8

Evaluating Breast Cancer Cell Proliferation, Colony Formation, and Migration

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MCF-7, LCC9 and LY2 cells were placed in hormone-depleted medium and transfected with precursor microRNA or anti-miR transcripts or 48 h and the following assays performed. For BrdU assays, cells were treated with 100 nM 4-OHT for 48 h and cell proliferation quantified using BrdU ELISA assay (Roche). For colony formation, following transfection, LY2 cells were counted and transferred to agarose plates and allowed to grow for 14 days to form colonies. Colonies were stained with crystal violet (0.005% w/v, Sigma) and counted using an inverted Nikon microscope (×10 objective). For cell migration assays, LY2 cells grown in hormonally depleted medium and transfected in six well plates for 24 h. Cells were wounded by scratching with a p20 pipette tip, and treated with 100 nM 4-OHT for up to 8 h. Images were captured at each time point at 4× magnification using an EVOS microscope (Thermo Fisher Scientific Waltham, MA, USA). NIH Image J software (http://imagej.nih.gov/ij/) was used to analyze percent wound area.
Muluhngwi P., Krishna A., Vittitow S.L., Napier J.T., Richardson K.M., Ellis M., Mott J.L, & Klinge C.M. (2016). Tamoxifen differentially regulates miR-29b-1 and miR-29a expression depending on endocrine-sensitivity in breast cancer cells. Cancer letters, 388, 230-238.
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9

Evaluation of cytotoxicity and proliferation

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Fetal bovine serum, penicillin/streptomycin, and Dulbecco’s modified Eagle’s medium-high glucose were purchased from Sigma-Aldrich GmbH. (Germany). Methanol (MeOH), n-hexane, chloroform (CHCl3), ethyl acetate (EtOAc), and n-butanol (BuOH) used in HPLC analysis and extraction were HPLC grade and purchased from Merck. LDH Cell Cytotoxicity Assay (Roche 04 744 926 001, Germany) and BrdU ELISA Assay (Cat. No. 11 647 229 001, Germany) were supplied from Roche.
ŞAHİN YAĞLIOĞLU A., GÜNEŞ GÜRBÜZ D., DÖLARSLAN M, & DEMİRTAŞ İ. (2021). First determination of anticancer, cytotoxic, and in silico ADME evaluation of secondary metabolites of endemic Astragalus leucothrix Freyn & Bornm. Turkish Journal of Chemistry, 46(1), 169-183.
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10

BrdU Cell Proliferation Assay

Cited 1 time
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BrdU ELISA assay (Roche, 116472290001) was performed, as we previously described [41 (link)].
Becker-Greene D., Li H., Perez-Cremades D., Wu W., Bestepe F., Ozdemir D., Niosi C.E., Aydogan C., Orgill D.P., Feinberg M.W, & Icli B. (2021). MiR-409-3p targets a MAP4K3-ZEB1-PLGF signaling axis and controls brown adipose tissue angiogenesis and insulin resistance. Cellular and molecular life sciences : CMLS, 78(23), 7663-7679.
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