Cell proliferation was measured by BrdU (5-bromo-2′-deoxyuridine, Aldrich-Sigma, Gillingham, UK) assay [28 ]. Cultured tissue slices were incubated in 20 µM BrdU-containing medium for 3 or 24 h prior to fixation. The fixed cultures were incubated with 1N HCl for 10 min on ice followed by a 10-min incubation with 2N HCl at room temperature and placed in an incubator at 37 °C for 20 min. Acidic conditions were neutralized by placing the slides in borate buffer (0.1 M, pH = 8.5) for 12 min. Slides were washed three times for 5 min each with PBS and 1% Triton 100-X and permeabilized in a solution containing PBS (1 M) with 1% Triton 100-X, glycine (1 M), and normal goat serum (5%) for 1 h. The slides were then incubated overnight with anti-BrdU mono-antibody (eBioscience, Oxford, UK) at a dilution of 1: 50 in PBS followed by DAPI staining of the nuclei.
Anti brdu mono antibody
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The Anti-BrdU mono-antibody is a laboratory tool used for the detection and quantification of bromodeoxyuridine (BrdU) incorporation into DNA. It can be used in various cell biology and molecular biology applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using anti brdu mono antibody
1
BrdU Proliferation Assay in Tissue Slices
2
Assessing Neuronal Mitotic Behavior with BrdU Kit
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A BrdU (5-bromo-2‘-deoxyuridine) Assay Kit (Aldrich-Sigma) was utilised to assess the mitotic behaviour of neuronal cells. In brief, tissue sections were incubated in a medium containing 20 µM BrdU for 24 h. Next, tissues were fixed and incubated with 1 N HCl on ice for 10 min, incubated with 2 N HCl at room temperature for 10 min, and then placed in an incubator at 37 °C for 20 min. To neutralise the acid, the tissues were incubated in 0.1 M borate buffer (pH of 8.5) at room temperature for 12 min, and then washed three times in a solution of PBS and 1% Triton 100-X. Following this, the tissues were permeabilised in a solution containing 1 M glycine, 5% normal goat serum, PBS, and 1% Triton 100-X for an hour, and then incubated overnight with anti-BrdU mono-antibody (eBioscience) in PBS (1:50). Finally, they were stained with DAPI and double immunostained to examine the phenotype of the new cells.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!