| The immunocytochemical detection of bromodeoxyuridine (BrdU) incorporated into DNA is a powerful tool to study the cytokinetics of normal and neoplastic cells. In vitro or in vivo labeling of cells with the thymidine analogue BrdU and the subsequent detection of incorporated BrdU with specific anti-BrdU monoclonal antibodies is an accurate and comprehensive method to quantitate the degree of DNA-synthesis. BrdU is incorporated into the newly synthezised DNA of the S-phase cells and can thus provide an estimate for the fraction of cells in S-phase. Also dynamic proliferative information (such as the S-phase transit rate and the potential doubling time) can be obtained, by means of bivariate BrdU/DNA flow cytometric analysis. |
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| Catalog # | B2850-01L |
| Applications | Suitable for use in Immunofluorescence, Flow Cytometry, Immunohistochemistry and Immunocytochemistry. Other applications not tested. |
| Recommended Dilution | Flow Cytometry/FACS: 1:25-1:200 |
| Immunocytochemistry: 1:300 |
| Immunofluorescence: 1:300 |
| Immunohistochemistry (FFPE): 1:300 |
| *Denaturation of the DNA is critical for successful staining of BrdU. This can be achieved by exposing cells to heat, or acid. For heat-induced epitope retrieval, 10mM citrate buffer pH 6.0 is recommended. Alternatively, a 30 min incubation in 2M HCl can be performed. The HCl must then be neutralized for 2 min with 0.1 M Na2B4O7. Pretreatment of tissues with proteinase K should be avoided. |
| Optimal dilutions to be determined by the researcher. |
| Storage and Stability | May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. |
| CAS Number | n/a |
| Clone Type | Monoclonal |
| Isotype | IgG2a |
| Clone No | 8J386 |
| Host | Rat |
| Concentration | ~1mg/ml |
| Form | Supplied as a liquid in PBS, 0.1% sodium azide, 50% glycerol. |
| Purity | Purified by Protein G affinity chromatography. |
| Immunogen | Bromodeoxyuridine (BrdU). Cellular Localization: Nuclear. |
| Specificity | Recognizes BrdU in single stranded DNA, as well as BrdU attached to a protein carrier or free BrdU. It detects nucleated cells in S-Phase which have had BrdU incorporated into their DNA. Also reacts with chlorodeoxyuridine but with reduced staining. Species crossreactivity: All species. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
