BrdU IHC Kit

Catalog No. ABIN955804

Product Details

Target: BrdU (Bromodeoxyuridine (BrdU))

Application: Immunohistochemistry (IHC)

Components: Trypsin Enzyme Conc., 4x: 3 mL
Trypsin Dilution Buffer: 12 mL
Denaturing Solution: 6 mL
Blocking Buffer: 6 mL
Detection Antibody (biotinylated & prediluted): 6 mL
Streptavidin-HRP Conjugate (prediluted): 6 mL
Substrate Reaction Buffer: 6 mL
DAB Concentrate: 0.3 mL
Hematoxylin Counterstain: 6 mL
Mounting Media: 6 mL
5 Control Slides: Intestinal tissue from mouse injected with BrdU. 1 x Positively stained, 4 x Unstained,
The material in this kit is sufficient to run 50 slides. The average test area is defined as a circle around the tissue with an approximate diameter of 2 cm.

Trypsin is only required if using formalin fixed tissues. If the tissues are fixed in alcohol, trypsin digestion is not required.

Material not included: Hydrogen peroxide (30% solution) for quenching endogenous peroxidase activity
Phosphate buffered saline (PBS) solution
Distilled water
Ethyl alcohol
Xylene
Coverslips
Bromodeoxyuridine (BrdU)
Methanol

Application Details

Restrictions: For Research Use only

Handling

Format: Liquid

Precaution of Use: Wearing of latex or rubber gloves is recommended when running this kit, especially avoid contact of DAB reagent with skin and clothes.

Storage: -20 °C

Storage Comment: Upon receipt, store the entire kit at -20°C. Once the kit is thawed, you may keep at 4°C for 5 days. For long term storage, it is recommended you aliquot and freeze the components at -20°C, particularly the Streptavidin-HRP Conjugate, the Detection Antibody and the Trypsin Concentrate.

Target Details

Target: BrdU (Bromodeoxyuridine (BrdU))

Alternative Name: BrdU (BrdU Products)

Target Type: Chemical

Background: A non-isotopic immunohistochemical staining for the localization of DNA synthesis and cell proliferation. Evaluation of cell cycle progression is essential for investigations in many scientific fields. Measurement of [ 3 H] thymidine incorporation as cells enter S phase has long been the traditional method for the detection of cell proliferation. Subsequent quantification of [ 3 H] thymidine is performed by scintillation counting or autoradiography. This technique is slow, labor-intensive and has several limitations including the handling and disposal of radioisotopes and the necessity of expensive equipment. A well-established alternative to [ 3 H] thymidine uptake has been demonstrated by numerous investigators. In these methods BrdU, a thymidine analog, replaces [ 3 H] thymidine. BrdU is incorporated into newly synthesized DNA strands of actively proliferating cells. Following partial denaturation of double stranded DNA, BrdU is detected immunochemically allowing the assessment of the population of cells which are actively synthesizing DNA. The BrdU IHC Kit involves incorporation of BrdU into proliferating cells, in vivo or in vitro, and visual staining (dark brown nuclei) of these cells which is achieved using a biotinylated anti-BrdU antibody followed by Streptavidin-HRP Conjugate and diaminobenzidine (DAB) substrate.