Polyclonal Antibody TO 5'-BROMO-2'DEOXYURIDINE
Polyclonal Antibody TO 5'-BROMO-2'DEOXYURIDINE
-For Immunoprecipitation, histochemistry, brdU incporation -broad mammalian species reactivity
PRODUCT CODE : RDI-BRDUabS $438.00/vial
SOURCE: Sheep sera
Immunogen: Highly purified bromodeoxyuridine
PURIFICATION METHOD: DEAE chromatography
SPECIFICITY: very low cross reactivity to 5-methyl cytosine (see notes below)
References:-Campana, D. et al, Dept of Immunology Royal Free Hospital School of Medicine, London U.K.
-Journal of Immunological Methods 107 (1):79-88, 1988 February 24.
PRESENTATION: delivered in a sealed vial at a concentration of 250µg in 0.25ml PBS buffer (no Azide).We recommend that each laboratory determine an optimum working titre for use in its particular application.
STORAGE: Purified antibodies are generally stable for long periods at 4oC. However repeated warming to room temperature and re-cooling may result in a loss of activity and hence effective working titre. It is recommended that monoclonal antibodies are aliquotted upon receipt and then stored at -20C. After thawing they should then be used as a stock solution for up to 60 days. Dilute working solutions are best prepared on the day of use.
For research use only. Not supplied for use in human diagnostic or therapeutic procedures
Background: Bromodeoxyuridine was coupled to keyhole limpet hemocyanin (KLH) and was used to immunize sheep. The sheep polyclonal antibody is provided as non-sterile antiserum purified. The antibody was tested using immunoprecipitation against 5-Methyl Cytosine (5-MeC) and bromodeoxyuridine (BrdU) or control (no anitgen) and assayed by A405 spectrophotometry. At a concentration of 25ug/ml, this product demonstartes 8 fold higher reactivity against BrdU verus 5-MeC. Antibody titers out at 50ng/ml. For best results, use the product at a concentration of 25 to 100ug/ml. Nearly complete immunoprecipitation of BrdU was obtained at a concentration of 100 to 500ug/ml. The product has also been tested for utility for staining of brdU incorporated into DNA of replicating cells. When utilized in 10 microgram/ml concentration, the product is comparable to commercially available monoclonal antibodies commonly used for the same purpose.
Immunohistochmeistry Protocol for anti-Bromodeoxyuridine:
-fixation in cold methanol for 30 minutes followed by immersion in 0.007N NaOH for 10-15 seconds allows Brdu staining with simultaneous detection of nuclear, cytoplasmic and membrane antigens as well as preservation of morphological detail. This method is optimal for detection of nuclear Ki67 and TdT. -original work performed Dr. David Stoller, Tufts University, Boston Ma.