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5 bromo 2 deoxyuridine brdu labeling reagent

Manufactured by Roche
Sourced in United States

5-Bromo-2′-deoxyuridine (BrdU) Labeling reagent is a synthetic thymidine analog that can be incorporated into the DNA of dividing cells during the S-phase of the cell cycle. The presence of BrdU can be detected using specific antibodies, allowing for the identification and quantification of proliferating cells.

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4 protocols using 5 bromo 2 deoxyuridine brdu labeling reagent

1

Cell Proliferation and Signaling Assays

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Sp was purchased from Sigma-Aldrich (St. Louis, MO, USA). 5-Bromo-2′-deoxyuridine (BrdU) Labeling reagent and the Detection Kit I were obtained from Roche Diagnostics GmbH (Mannheim, Germany). Anti-cyclin D1 and anti-p27 antibodies were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Anti-mitogen-activated protein kinase (MAPK), anti-PI3K p85 and anti-AKT antibodies were all purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Alkaline phosphatase-labeled goat anti-rabbit IgG antibody and goat anti-mouse IgG antibody were both from Sigma-Aldrich.
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2

Measuring Osteoblast and Osteocyte Proliferation

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To detect the extent of cell proliferation in the osteoblast and osteocyte populations, 5′-bromo-2′deoxyuridine (BrdU) labeling reagent (45 μg g−1 body weight; Roche, Indianapolis, IN, USA) was injected intraperitoneally into mice at P10. Two hours after injection, their femora were dissected, embedded and sectioned in the mid-sagittal plane for immunodetection with a BrdU labeling and detection kit (Roche). For statistical analyses, three independent littermates were used in each of the studies.
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3

Evaluation of Mesangial Cell Proliferation

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Primary human mesangial cells were grown to near confluence in 96-well culture plates (Nunc) using CS-C complete serum free medium and kept in resting condition as described above. Cells were left unstimulated or incubated with stimulants suspended in CS-C serum-free-maintenance medium containing 5-bromo-2′-deoxyuridine (BrdU)-labeling reagent (100 μM, Roche Diagnostics) for 24 h. Stimulants included galactose-deficient polymeric IgA1-IgAN (100 μg/mL), M4 or M4Δ451 (both at 10 μg/mL) as well as M4 and M4Δ451 pre-incubated for 1 h with galactose-deficient polymeric IgA1-IgAN. The positive control was murine platelet-derived growth factor (PDGF)-BB (50 ng/ml, PeproTech Inc, Rocky Hill, NJ). Proliferation was detected using a calorimetric BrdU cell proliferation ELISA-kit (Roche Diagnostics) according to manufacturer's instructions.
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4

Detecting Cell Proliferation in Developing Roots

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To detect cell proliferation in developing roots, 5'-bromo-2' deoxyuridine (BrdU) labeling reagent (45 µg/g body weight; Roche, Indianapolis, IN, USA) was injected intraperitoneally into 10-day-old wild type (WT) and mutant (MT) mice. Two hours after injection, mice were dissected, the mandibles were fixed with Carnoy's fixatives at 4℃ overnight, embedded in paraffin, and sectioned in the root elongation region and furcation region for immunodetection with the BrdU labeling and detection kit (Roche). For statistical analysis, 5 consecutive sections from 3 independent littermates were used.
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