Paclitaxel was purchased from Calbiochem, Propidium iodide, methanol, DMSO and 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) reagents were procured from Sigma Aldrich. DeadEnd Fluorometric TUNEL system was from Promega. JC1dye-5,5′,6,′-tetrachloro-1, 1′, 3, 3′ tetraethylbenzimidazolylcarbocyanine iodide was from Molecular Probes Inc., Eugene, USA. Enhanced chemiluminescence (ECL) reagent and antibodies were from Santa Cruz, USA while the VEGF quantikine ELISA kit was from R& D system (Minneapolis, USA). Dulbecco-modified Eagle’s medium (DMEM), fetal bovine serum (FBS), streptomycin and penicillin were from Gibco-BRL, USA. Lipofectamine 2000 was from Invitrogen, USA while the BrdU labeling kit was from Roche Diagnostics, Indianapolis, USA.
Brdu labeling kit
Manufactured by Roche
Sourced in United States
The BrdU labeling kit is a laboratory product designed to detect cell proliferation. It allows for the incorporation of bromodeoxyuridine (BrdU) into newly synthesized DNA, which can then be detected using specific antibodies. The kit provides the necessary reagents and protocols to perform this analysis.
Automatically generated - may contain errors
Lab products found in correlation
N2 supplement, by Thermo Fisher Scientific (2 mentions)
Cell counting kit 8, by Dojindo Laboratories (2 mentions)
Vegf quantikine elisa kit, by R&D Systems (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Deadend fluorometric tunel system, by Promega (1 mentions)
Axio imager, by Zeiss (1 mentions)
Imark microplate reader, by Bio-Rad (1 mentions)
15 protocols using brdu labeling kit
1
Cytotoxic Effects of Paclitaxel on Cancer Cells
2
Evaluating Cell Viability and Proliferation
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Cell viability was determined by Alamar blue assay. Briefly, cells were plated into a 96-well plate at the concentration of 3 × 104/mL. At different time point after Adv-MMP-10 infection, AlamarBlue reagent was introduced to each well at a volume of 10% of total well volume, and allowed to incubate for 2 h. Absorbance readings at wavelengths of 570 nm and 600 nm were compared to the absorbance of negative control wells containing media and AlamarBlue only.
Cell proliferation was determined using a BrdU labeling kit (Roche, Indianapolis, IN, USA). The BrdU incorporation was detected by FITC-BrdU antibody and analyzed under a fluorescent microscope. The experiments were performed in triplicate.
Cell proliferation was determined using a BrdU labeling kit (Roche, Indianapolis, IN, USA). The BrdU incorporation was detected by FITC-BrdU antibody and analyzed under a fluorescent microscope. The experiments were performed in triplicate.
3
Proliferation Assay with BrdU
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Cells were plated on coverslips and treated as described above. Cells proliferation was determined using a BrdU labeling kit (Roche, Indianapolis, IN, USA). The BrdU incorporation was detected by FITC-BrdU antibody (Roche) and analyzed under a fluorescent microscope. The experiments were performed in triplicate.
4
Derivation of Neural Progenitor Cells from hESCs
5
Cell Proliferation by BrdU Quantification
6
Senescence-Associated Beta-Galactosidase and BrdU Assay
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We detected SA-β-Gal as described (Dimri et al., 1995 (link)), using a commercial kit (BioVision). Briefly, 30,000 cells per well were seeded on a 12-well plate. Cells were fixed using 500 μL Fixing Buffer for 10 min, followed by three washes in PBS, and overnight incubation at 37°C in Staining Solution. Cells were visualized and counted using an inverted microscope.
DNA synthesis was determined using a modified BrdU labeling kit (Roche #11296736001). Cells were cultured in BrdU (20 μM in DMSO) for 24 hours, fixed in 4% neutral-buffered formalin for 10 min, and washed 3 times in ice cold PBS. Permeabilization was perform for 30 min by incubation in 0.5% Triton X-100 in PBS. Nuclear DNA was digested by a combination of DNase I and exonuclease III for 1 hour, and BrdU-containing fragments were visualized by immunofluorescence.
DNA synthesis was determined using a modified BrdU labeling kit (Roche #11296736001). Cells were cultured in BrdU (20 μM in DMSO) for 24 hours, fixed in 4% neutral-buffered formalin for 10 min, and washed 3 times in ice cold PBS. Permeabilization was perform for 30 min by incubation in 0.5% Triton X-100 in PBS. Nuclear DNA was digested by a combination of DNase I and exonuclease III for 1 hour, and BrdU-containing fragments were visualized by immunofluorescence.
7
Senescence-Associated Beta-Galactosidase and BrdU Assay
8
Npnt Overexpression Promotes M3H1 Cell Proliferation
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M3H1 cells were transfected with an Npnt expression vector, then cultured in Npnt-coated dishes for 48 hours. Proliferation was determined using a Cell Counting Kit (CCK)-8 (Dojindo Laboratories), according to the manufacturer’s protocol, while BrdU incorporation was assayed using a BrdU labeling kit (Roche Diagnostics). After 48 hours of culture, BrdU was applied to the plates for 30 minutes, then incorporated BrdU was detected after washing with PBS, according to the manufacturer’s protocol. BrdU-positive cells were counted under a fluorescence microscope.
9
Quantifying Cell Proliferation Dynamics
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Cells were plated in 8-chamber slides and treated with the indicated concentrations of drugs for 24h followed by labeling with BrdU for 1h at 37°C and analyzed using the BrdU labeling kit from Roche (Indianapolis, IN), following the manufacturer's protocol. BrdU stained slides were mounted using Fluro-Gel and analyzed using Zeiss AxioImager for brightfield microscopy. Proliferation in response to treatment was measured as percent BrdU positive cells from the total number of observed cells in the chamber.
10
Pkp1 siRNA Impacts CLDE Cell Proliferation
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CLDE cells were cultured in 96-well plates at 0.8 × 104 cells/well for 72 h, then transfected with control or Pkp1 siRNA. Proliferation was determined using a Cell Counting Kit (CCK)-8 (Dojindo Laboratories, Kumamoto, Japan) according to the manufacturer’s protocol. Briefly, 10 μl of CCK-8 solution was added to each well and optical density was measured at 450 nm using an iMark microplate reader (Bio-Rad). Cell proliferation was also assayed by BrdU incorporation using a BrdU labeling kit (Roche Diagnostics, Indianapolis, IN, USA). Cells transfected with control or Pkp1 siRNA were cultured for 48 h. BrdU was added to the plates for 30 min, then incorporated BrdU was detected after 3 washes in PBS according to the manufacturer’s protocol. BrdU-positive cells were counted and the incorporation ratio was calculated by visualizing the cells with a confocal microscope.
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