This BrdU Immunohistochemistry BioAssay™ Kit is a histochemical staining kit for the detection and localization of bromodeoxyuridine incorporated into newly synthesized DNA of actively proliferating cells.
Kit Features:More efficient animal-friendly manufacturing processThe MCF-7 cells are labeled with an exact concentration of BrdU, meaning low/negligible batch-to batch variationControls are more relevant to experimental samples. Many researchers and pharmaceutical companies use the BrdU kit to test the efficacy of drugs that influence proliferative capacity using specific cell culturesSlides can be kept frozen while still maintaining their morphology, and can withstand multiple freeze-thaw cycles Ease of interpretation compared to tissue sections, no knowledge of the morphology of intestinal tissue is required (see sample pictures)
Applications:Immunohistochemistry on formalin fixed, paraffin embedded, as well as frozen tissue sections. Furthermore, this kit can be used for the immunocytochemical staining of cultured cells on glass slides or cell culture plates. Optimal concentration of the primary antibody should be evaluated by serial dilutions.
Test Principle:A non-isotopic immunohistochemical staining procedure for the localization and quantification of DNA synthesis and cell proliferation. Evaluation of cell cycle progression is essential for investigations in many scientific fields. Measurement of [3H] thymidine incorporation as cells enter S phase has long been the traditional method for the detection of cell proliferation
Subsequent quantification of [3H] thymidine is performed by scintillation counting or autoradiography. This technology 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 [3H] thymidine uptake has been demonstrated by numerous investigators. In these methods bromodeoxyuridine (BrdU), a thymidine analog, 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.
This BrdU Immunohistochemistry BioAssay™ 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 Prediluted Biotinylated Sheep anti-BrdU Detector Antibody followed by Streptavidin-HRP Conjugate and DAB (diaminobenzidine) substrate.
Storage and Stability:Upon receipt, store entire kit at -20ºC. Thawed kit may be kept at 4ºC for 5 days. For long-term storage, it is recommended to aliquot and freeze the components at –20ºC, particularly the Streptavidin-HRP Conjugate, the Prediluted Biotinylated Sheep anti-BrdU Detector Antibody, the 4x Trypsin Concentrate and the control slides. Wearing of latex or rubber gloves, a lab coat, and safety goggles is recommended when running this kit; especially avoid inhalation of DAB reagent and contact with skin and clothes.
