Brdu cell proliferation kit

Manufactured by Merck Group

Sourced in United States, Germany

The BrdU Cell Proliferation Kit is a laboratory tool used to measure cell proliferation. It utilizes the incorporation of the nucleoside analogue bromodeoxyuridine (BrdU) into the DNA of dividing cells, allowing for the quantification of cell proliferation.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Cell Proliferation Kit (18 citations)

34 protocols using brdu cell proliferation kit

Generation and Maturation of Dendritic Cells for T Cell Proliferation Assay

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

CD14⁺ monocytes and naïve CD4⁺ T cells were purified from peripheral blood mononuclear cells (PBMCs) obtained from healthy consenting donors, as reported in the previous study.¹² DCs were generated by culturing CD14⁺ monocytes in RPMI 1640 medium containing 10% FBS (Invitrogen, Carlsbad, CA), 20 ng/mL GM-CSF, and 10 ng/mL IL-4 (R&D Systems, Minneapolis, MN) for 5 days. The medium was replaced with fresh medium containing GM-CSF and IL-4 on day 3. For maturation of DCs, immature DCs were stimulated with LPS (100 ng/ml) after priming with interferon-γ (IFN-γ) for 3 h. CL1-5 and A549-LCAF conditioned DCs were generated by culturing CD14⁺ monocytes in RPMI 1640 medium containing FBS, GM-CSF, and IL-4 presenting in CL1-5-LCAF-CM or A549-LCAF-CM (20%), then stimulated as described above.
MLR was carried out by culturing naïve CD4⁺ T cells for a set number of mature DCs (10⁴cells/well) in 96-well plates for 5 days. Human T cells were purified from PBMCs obtained from healthy consenting donors using immunomagnetic CD4 naive T cell MACS beads and a MACS LS column (Miltenyi Biotec Inc, Auburn, CA). The T cell proliferation was assessed by BrdU Cell Proliferation Kit (EMD Millipore, Billerica, MA), as described in a previous study [14 (link)].

Hsu Y.L., Hung J.Y., Chiang S.Y., Jian S.F., Wu C.Y., Lin Y.S., Tsai Y.M., Chou S.H., Tsai M.J, & Kuo P.L. (2016). Lung cancer-derived galectin-1 contributes to cancer associated fibroblast-mediated cancer progression and immune suppression through TDO2/kynurenine axis. Oncotarget, 7(19), 27584-27598.

+ Open protocol

+ Expand

MEFs Proliferation Assay with BrdU

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

MEFs were plated on 96-well plates and incubated for 24 h. The cells were labeled with BrdU for 2 h, and the incorporation of BrdU was analyzed using a BrdU cell proliferation kit (Merck Millipore, Darmstadt, Germany) according to the manufacturer's protocol. Each value was normalized by cell number using a Cell Counting Kit-8.

Yamazaki H., Kasai S., Mimura J., Ye P., Inose-Maruyama A., Tanji K., Wakabayashi K., Mizuno S., Sugiyama F., Takahashi S., Sato T., Ozaki T., Cavener D.R., Yamamoto M, & Itoh K. (2020). Ribosome binding protein GCN1 regulates the cell cycle and cell proliferation and is essential for the embryonic development of mice. PLoS Genetics, 16(4), e1008693.

+ Open protocol

+ Expand

Effects of HA and 4-MU on MG Cell Proliferation

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

The effect of HA on MG cell proliferation was assayed in vitro by measuring the incorporation of 5-bromo-2-deoxyuridine (BrdU) into the newly synthesized DNA of replicating cells (BrdU Cell Proliferation Kit, EMD Millipore, Darmstadt, Germany). The hMGCs were plated at a seeding density of 5000 cells/well in 96-well plates and allowed to grow for 24 hours in the presence or absence of HA or 4-methylumbelliferone (4-MU; Sigma-Aldrich Corp.). The concentrations used were 0.02% and 0.05% of HMWHA; 0.02% and 0.05% of LMWHA; or 0.25 mM, 0.5 mM, or 2 mM of 4-MU. After 24 hours, with the cells in their exponential growth phase, BrdU was added to each well and left for 2 hours. The cells were then fixed and the detection of BrdU incorporation carried out according to the manufacturers' instructions. Absorbance in each well was read as optical density (OD) by using a spectrophotometer microplate reader (Fluostar Omega, BMG Labtech, Offenburg, Germany) set at a test wavelength of 450 nm. Each condition was tested in triplicate and three separate experiments were carried out. The mean of each individual experiment is presented as a single point on the graph.

Sun M., Puri S., Parfitt G.J., Mutoji N, & Coulson-Thomas V.J. (2018). Hyaluronan Regulates Eyelid and Meibomian Gland Morphogenesis. Investigative Ophthalmology & Visual Science, 59(8), 3713-3727.

+ Open protocol

+ Expand

Cell Proliferation Assay with BrdU

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

Approximately 8,000 cells were plated per well on 96-well plates and incubated overnight at 37°C. The cells were treated with B6H12 or isotype control antibody (1 μg/ml) for 24 h. BrdU was added for 4 or 24 h as indicated in the figures, and BrdU incorporation was quantified using a BrdU Cell Proliferation Kit according to the manufacturer's instructions (EMD Millipore). For flow analysis, MDA-MB-231 cells were labeled with BrdU, and unlabeled cells were used as a negative control.

Kaur S., Elkahloun A.G., Singh S.P., Chen Q.R., Meerzaman D.M., Song T., Manu N., Wu W., Mannan P., Garfield S.H, & Roberts D.D. (2016). A function-blocking CD47 antibody suppresses stem cell and EGF signaling in triple-negative breast cancer. Oncotarget, 7(9), 10133-10152.

+ Open protocol

+ Expand

Medulloblastoma Cell Proliferation Assay

Cited 1 time

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

Medulloblastoma cells were obtained from medulloblastoma allografts by mechanical dissociation and viable cell fractioning. The cells were maintained in Neurobasal A medium (Invitrogen) containing B-27 supplement (Invitrogen), epidermal growth factor 20 ng/ml (Invitrogen), basic fibroblastic growth factor 20 ng/ml (Invitrogen), nonessential amino acids (Invitrogen), N-acetyl cysteine 60 mg/ml, and Glutamax (Invitrogen) [20 (link)].
Medulloblastoma cells were seeded into 96-well plates, followed by treatment with various concentrations of BBR for 36 h. The Brdu assays were conducted using the Brdu Cell Proliferation Kit (Merck Millipore; Bedford, MA) according to manufacturer’s instructions.

Wang J., Peng Y., Liu Y., Yang J., Ding N, & Tan W. (2015). Berberine, a natural compound, suppresses Hedgehog signaling pathway activity and cancer growth. BMC Cancer, 15, 595.

+ Open protocol

+ Expand

Quantifying Cell Proliferation with BrdU

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

The BrdU incorporation was measured using the BrdU Cell Proliferation Kit (Merck, Germany). The cells were seeded in a 96 well plate at a concentration of 0.8x10⁵ cells/ml and left overnight. The next day, the cells were treated with three different concentrations of flavokawain B. 24 h before fixing the cells, 20 μl of BrdU was added to each well. After the respective incubation hours, the cells were fixed and denatured using a fixing solution. The plates were then stored at 4 °C. Then, the plates were washed twice before adding 100 μl of the detector antibody into each well for 1 h. Next, 100 μl of Goat anti-mouse Ig G-HRP conjugated was added for 30 min. Afterwards, the plates were incubated with 100 μl of the TMB substrate for roughly 30 min. Finally, 100 μl of stop solution was added and the absorbance was measured at 450 nm, using the μquant plate reader (Bio-tek Instruments, USA).

Abu N., Akhtar M.N., Yeap S.K., Lim K.L., Ho W.Y., Abdullah M.P., Ho C.L., Omar A.R., Ismail J, & Alitheen N.B. (2016). Flavokawain B induced cytotoxicity in two breast cancer cell lines, MCF-7 and MDA-MB231 and inhibited the metastatic potential of MDA-MB231 via the regulation of several tyrosine kinases In vitro. BMC Complementary and Alternative Medicine, 16, 86.

+ Open protocol

+ Expand

NIH-3T3 Cell Proliferation Assay

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

NIH-3T3 cells (ATCC, Manassas, VA) were maintained with DMEM containing 10% FCS and 0.1% β-mercaptoethanol, and cultured with reduced serum media (1% FCS) overnight prior to use. A total of 5×10³ cells were cultured with 200 μl complete media containing rPAI-1₂₃ (0, 0.6 and 10 nM). Cell proliferation was assessed using a BrdU cell proliferation kit (EMD Millipore, Billerica, MA), per manufacturer's instructions.

Chung E.J., McKay-Corkum G., Chung S., White A., Scroggins B.T., Mitchell J.B., Mulligan-Kehoe M.J, & Citrin D. (2015). A truncated Plasminogen Activator Inhibitor-1 protein protects from pulmonary fibrosis mediated by irradiation in a murine model. International journal of radiation oncology, biology, physics, 94(5), 1163-1172.

+ Open protocol

+ Expand

Assessing Fibroblast and Pneumocyte Proliferation

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

Enriched pneumocytes, primary fibroblasts and NIH-3T3 (ATCC®, Manassas, VA), a murine fibroblast cell line, were maintained with DMEM containing 10% FBS and 0.1% β-mercaptoethanol. NIH-3T3 and primary cells were cultured with DMEM containing 1% FBS overnight prior to use in proliferation assays. To assess the proliferation of fibroblasts and pneumocytes in response to TGF-α, 5 × 10³ cells were cultured with 200 ll complete media containing TGF-α (0.1, 1, 10 ng/mL). Cell proliferation was assessed using a BrdU cell proliferation kit (EMD Millipore) following the manufacturer’s instruction.

Chung E.J., Hudak K., Horton J.A., White A., Scroggins B.T., Vaswani S, & Citrin D. (2014). Transforming Growth Factor Alpha is a Critical Mediator of Radiation Lung Injury. Radiation research, 182(3), 350-362.

+ Open protocol

+ Expand

BrdU Proliferation Assay of 4T1 Cells

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

The BrdU proliferation assay was measured using a BrdU cell proliferation kit (Merck, Germany). Firstly, the 4T1 cells (8 × 10⁴ cells/mL) were seeded in the plates and incubated overnight. The next day, the cells were treated with two different concentrations (IC₅₀=0.27 mg/mL; IC₇₅=0.35 mg/mL) of coconut juice vinegar, added with 20 μL of BrdU and incubated for another 72 h. Later, the cells were fixed and denatured using a fixing solution and stored at 4°C. The plates were washed and then incubated with 100 μL of the detector antibody per well for 1 h. Next, 100 μL of goat anti-mouse immunoglobulin G-horseradish peroxidase (HRP) conjugate was added for 30 min, followed by further incubation with 100 μL of the 3,3′,5,5′-tetramethylbenzidine substrate for 30 min. Finally, 100 μL of stop solution was added to the plates, and the absorbance was measured at 450 nm, using a μQuant plate reader (Bio-Tek Instruments, Vermont, USA).

Mohamad N.E., Yeap S.K., Abu N., Lim K.L., Zamberi N.R., Nordin N., Sharifuddin S.A., Long K, & Alitheen N.B. (2019). In vitro and in vivo antitumour effects of coconut water vinegar on 4T1 breast cancer cells. Food & Nutrition Research, 63, 10.29219/fnr.v63.1616.

+ Open protocol

+ Expand

BrdU Cell Proliferation Assay Protocol

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

Cell proliferation assay was performed using the BrdU Cell Proliferation Kit (#2750, EMD Millipore). Equal quantities of cells were seeded into 96-well plates. Following attachment, cells were incubated in BrdU for 4 hrs, and BrdU incorporation was measured spectrophotometrically as per kit instructions using TMB as a substrate. Absorbance was recorded at 450 nm. Each experiment was performed at least in triplicate.

Ogden A., Garlapati C., Li X.(., Turaga R.C., Oprea-Ilies G., Wright N., Bhattarai S., Mittal K., Wetherilt C.S., Krishnamurti U., Reid M.D., Jones M., Gupta M., Osan R., Pattni S., Riaz A., Klimov S., Rao A., Cantuaria G., Rida P.C, & Aneja R. (2017). Multi-institutional study of nuclear KIFC1 as a biomarker of poor prognosis in African American women with triple-negative breast cancer. Scientific Reports, 7, 42289.

+ 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!