In the past, most cell-proliferation studies have used radioactive thymidine as an incorporated label to give evidence of DNA replication. Recently, a less hazardous technique has been developed using bromodeoxyuridine (BrdU), a thymidine analog, in lieu of a radioactive reagent. (1) BrdU is incorporated into proliferating cells (S-phase) much in the same way as radioactive thymidine, but is then detected by a monoclonal anti-BrdU antibody, and revealed using a highly sensitive streptavidin- biotin staining system. BrdU has proven useful for proliferative studies of normal and neoplastic tissues both in vivo and in vitro.
The BrdU staining system uses a biotinylated monoclonal anti-BrdU, thus eliminating the need for a species-specific secondary antibody. As a result, BrdU labeling can be performed in rats and mice without problems of crossreactivity or spurious staining.
Streptavidin-peroxidase is used as a signal generator for the BrdU system. Diaminobenzidine (DAB) in the presence of hydrogen peroxide is used as a chromogen, staining BrdU-incorporated nuclei dark brown.
This kit also contains four BrdU control slides for the convenience of the investigator: four unstained positive controls.
B2851-01 Trypsin Concentrate 1x3ml
B2851-02 Trypsin Diluent 1x12ml
B2851-03 Denaturing Solution 1x6ml (Ready-to-use)
B2851-04 Blocking Solution 1x6ml(Ready-to-use)
B2851-05 BrdU (Biotin) 1x6ml (Ready-to-use)
B2851-06 Streptavidin (HRP) 1x6ml (Ready-to-use)
B2851-07 Substrate Buffer, 20x 1x2ml
B2851-08 DAB, 20x 1x2ml
B2851-09 0.6% Hydrogen Peroxide, 20x 1x2ml
B2851-10 Hematoxylin 1x6ml (Ready-to-use)
B2851-11 Histomount(TM) 1x6ml (Ready-to-use)
B2851-12 Control Slides 4x Unstained Controls
Materials Required but not Provided:
-Phosphate Buffered Saline (PBS)
-Quenching solution for endogenous peroxidase
-Alcohol and xylene
Storage and Stability:
Store at 4 degrees C. Stable for at least 6 months.