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Rabbit anti brdu

Manufactured by Merck Group

Rabbit anti-BrdU is a laboratory reagent used to detect and quantify proliferating cells. It is a polyclonal antibody that specifically binds to bromodeoxyuridine (BrdU), a synthetic thymidine analog that is incorporated into the DNA of dividing cells during the S-phase of the cell cycle. This antibody can be used in a variety of cell-based assays, such as immunohistochemistry and flow cytometry, to identify and measure the proportion of cells undergoing DNA replication and cell division.

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2 protocols using rabbit anti brdu

1

Quantification of Cumulus Cell Proliferation

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Cumulus cell proliferation was assessed by BrdU incorporation. Mice were injected with 100 mg BrdU/kg body weight 2 h prior to ovary collection. COCs were collected and fixed for 1 h in 4% paraformaldehyde. After 3 washes with PBS containing 0.1% Tween-20, COCs were permeabilized with 0.5% Triton X-100 for 20 min and blocked with 3% BSA in PBS for 30 min, then the COCs were incubated overnight with Rabbit anti-BrdU (100-fold dilution; Sigma) primary antibody. After thorough washing, the COCs were incubated for 1 h with Alexa Fluor 594-conjugated donkey anti-Rabbit secondary antibody (200-fold dilution; Thermo Fisher Scientific). The percentage of BrdU positive cumulus cells in relation to DAPI positive cells was determined by a confocal scanning laser microscopy (Nikon A1R-si) and the NIS Elements Basic Research software.
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2

Quantifying Pancreatic β-cell Proliferation

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The pancreas were fixed in 4% paraformaldehyde at 4°C overnight. The tissues were dehydrated, embedded in paraffin, and sectioned at 5 μm thickness. The primary antibodies used were as follows: rabbit anti-BrdU (1:50, Sigma-Aldrich), guinea pig anti-insulin (1:800, Dako), mouse anti-SFRP5 (1:50, Santa Cruz Biotech), and rabbit anti-glucagon (1:100, Santa Cruz Biotech). Incubations were performed overnight in a humidified chamber at 4°C. For BrdU/insulin staining, sections were treated with 1 M HCl at 37°C for 60 min before incubation with the primary antibody. Secondary antibodies were used as follows: Alexa Fluor 488 donkey anti-guinea pig (1:500, Jackson Immunoresearch), Alexa Fluor 488 donkey anti-rabbit (1:500, Jackson Immunoresearch), and Alexa Fluor 594 donkey anti-mouse (1:500, Life Technologies). Images were captured using an Olympus Microscope system.
After immunelabeling with BrdU and insulin, pancreatic islets in paraffin-embedded sections were photographed at ×400 and assigned blinded filenames. The number of BrdU (+) β-cells was manually counted. At least 2000 β-cells were counted per animal using three sections that were separated by at least 100 μm. Three animals per group were analyzed.
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