Technical Data
B2850-07X
Bromodeoxyuridine (BrdU) (BSA & Azide Free)
Description:
Bromodeoxyuridine (BrdU) is a thymidine analog and is specifically incorporated into DNA during DNA synthesis. 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 tumor 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.

Applications:
Suitable for use in Immunofluorescence, Flow Cytometry and Immunohistochemistry. Other applications not tested.

Recommended Dilution:
Immunohistochemistry(frozen & formalin/paraf n):1ug/ml for 30 minutes at RT.
For staining of formalin- xed tissues, incubate sections in 4N HCl for 30 minutes at RT, followed by digestion with trypsin at 1mg/ml PBS, 10 min at 37ΊC

Optimal dilutions to be determined by the researcher.

Positive Control:
Cultured cells grown in presence of BrdU or tissues from experimental animals injected with BrdU.

Cellular Localization:
Nuclear

Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20°C. Aliquots are stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
MabIgG10.N.35Affinity Purified
SizeStorageShippingSourceHost
100ug4°C (-20°C Glycerol)Blue IceMouse
Concentration:
~1mg/ml
Immunogen:
Bromodeoxyuridine (BrdU) labeled with BSA.
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as a liquid in 10mM PBS, pH 7.4 Also available with BSA and azide. See B2850-07.
Specificity:
Recognizes free BrdU, BrdU coupled to protein carrier, BrdU in ssDNA (partially denatured dsDNA). Species Reactivity: All species
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Bicknell S; van Eeden S; Hayashi S; Hards J; English D; Hogg JC. A non-radioisotopic method for tracing neutrophils in vivo using 5'-bromo-2'-deoxyuridine. American Journal of Respiratory Cell and Molecular Biology, 1994, 10(1):16–23. 2. Jones HB; Harbottle SJ; Bowdler AL. Assessment of the labeling index of cohorts of the anterior pituitary cell population in phenobarbital-treated male rats by a double immunohistochemical technique for bromodeoxyuridine and pituitary hormones. Journal of Histochemistry and Cytochemistry, 1994, 42(4):543–9. 3. Lloveras B; Garin-Chesa P; Myc A; Melamed M. In vitro bromodeoxyuridine labeling of malignant neoplasms. A comparative study with ow cytometry cell-cycle analysis. American Journal of Clinical Pathology, 1994, 101(6):703–7. 4. Williamson K; Gilliland R; Weir H; Grimes J; Hamilton P; Anderson N; Crockard A; Rowlands B. Hydrochloric acid denaturation of colorectal tumour tissue in ltrated with bromodeoxyuridine. Cytometry, 1994, 15(2):162–8. 5. Wilson MS; Scho eld PF. Correlation of PCNA with bromodeoxyuridine [letter]. Gut, 1994, 35(5):717–8. 6. Yamamura S; Okadome K; Onohara T; Komori K; Sugimachi K. Blood ow and kinetics of smooth muscle cell proliferation in canine autogenous vein grafts: in vivo BrdU incorporation. Journal of Surgical Research, 1994, 56(2):155–61. 7. Danova M; Pellicciari C; Zibera C; Mangiarotti R; Gibelli N; Giordano M; Wang E; Mazzini G; Riccardi A. Cell cycle kinetic effects of tamoxifen on human breast cancer cells. Flow cytometric analyses of DNA content, BrdU labeling, Ki-67, PCNA, and statin expression. Annals of the New York Academy of Sciences, 1993, 698:174–81. 8. Going JJ; Stanton PD; Cooke TG. In uences on measurement of cellular proliferation by histology and ow cytometry in mammary carcinomas labeled in vivo with bromodeoxyuridine. American Journal of Clinical Pathology, 1993, 100(3):218–22. 9. Hirano T; Zitsch RP 3d; Gluckman JL. Cell kinetics study of upper aerodigestive tract squamous cell carcinoma using bromodeoxyuridine. Annals of Otology, Rhinology and Laryngology,1993,102:42–6. 10. Jones HB; Clarke NA; Barrass NC. Phenobarbital-induced hepatocellular proliferation: anti-bromodeoxyuridine and anti-proliferating cell nuclear antigen immunocytochemistry. Journal of Histochemistry and Cytochemistry, 1993, 41:21–7. 11. Meyer JS; Koehm SL; Hughes JM; Higa E; Wittliff JL; Lagos JA; Manes JL. Bromodeoxyuridine labeling for S-phase measurement in breast carcinoma. Cancer, 1993, 71(11):3531–40. 12. Miwa H; Wada R; Abe H; Ohkura R; Yang SW; Watanabe H; Ogihara T; Hamada T; Sato N. Diagnosis of gastric adenoma versus early gastric cancer by bromodeoxyuridine immunohistochemistry from gastric biopsy specimen. Journal of Gastroenterology and Hepatology, 1993, 8(2):133–7. 13. Mollgard K; Schumacher U. Immunohistochemical assessment of cellular proliferation in the developing human CNS using formalin- xed paraf n-embedded material. Journal of Neuroscience Methods, 1993, 46(3):191–6. 14. Mourad WA; Connelly JH; Sembera DL; Atkinson EN; Bruner JM. The correlation of two argyrophilic nucleolar organizer region counting methods with bromodeoxyuridine-labeling index: a study of metastatic tumors of the brain [see comments]. Human Pathology, 1993, 24(2):206–10. 15. Patel S; Rew DA; Taylor I; Potten CS; Owen C; Roberts SA. Study of the proliferation in human gastric mucosa after in vivo bromodeoxyuridine labelling. Gut, 1993, 34(7):893–6. 16. Risio M; Candelaresi G; Rossini FP. Bromodeoxyuridine uptake and proliferating cell nuclear antigen expression throughout the colorectal tumor sequence. Cancer Epidemiology, Biomarkers and Prevention, 1993, 2(4):363–7. 17. Sanders EJ; Varedi M; French AS. Cell proliferation in the gastrulating chick embryo: a study using BrdU incorporation and PCNA localization. Development, 1993, 118(2):389–99. 18. Sebo TJ; Roche PC; Witzig TE; Kurtin PJ. Proliferative activity in non-Hodgkin’s lymphomas. A comparison of the bromodeoxyuridine labeling index with PCNA immunostaining and quantitative image analysis. American Journal of Clinical Pathology, 1993, 99(6):668–72. 19. Shibuya M; Ito S; Davis RL; Hoshino T. Immunohistochemical double staining with immunogold-silver and alkaline phosphatase to identify nuclear markers of cellular proliferation. Biotechnic and Histochemistry, 1993, 68(1):17–9. 20. Soriano E; Del Rio JA; Auladell C. Characterization of the phenotype and birthdates of pyknotic dead cells in the nervous system by a combination of DNA staining and immunohistochemistry for 5'-bromodeoxyuridine and neural antigens. Journal of Histochemistry and Cytochemistry, 1993, 41(6):819–27. 21. Yao CH; Kitazawa S; Fujimori T; Maeda S. In situ hybridization at the electron microscopic level using a bromodeoxyuridine labeled DNA probe. Biotechnic and Histochemistry, 1993, 68(3):169–74. 22. Biffo S; Verdun di Cantogno L; Fasolo A. Double labeling with non-isotopic in situ hybridization and BrdU immunohistochemistry: calmodulin (CaM) mRNA expression in post-mitotic neurons of the olfactory system. Journal of Histochemistry and Cytochemistry, 1992, 40:535–40. 23. Brito MJ; Filipe MI; Morris RW. Cell proliferation study on gastric carcinoma and non-involved gastric mucosa using a bromodeoxyuridine (BrdU) labelling technique. European Journal of Cancer Prevention, 1992, 1(6):429–35. 24. Dinjens WN; ten Kate J; Lenders MH; van der Linden EP; Bosman FT. Bromodeoxyuridine (BrdU) immunocytochemistry by exonuclease III (Exo III) digestion. Histochemistry, 1992, 98(3):199–205. 25. Ito S; Hoshino T; Shibuya M; Prados MD; Edwards MS; Davis RL. Proliferative characteristics of juvenile pilocytic astrocytomas determined by bromodeoxyuridine labeling. Neurosurgery, 1992, 31(3):413–8; discussion 419. 26. Lazarous DF; Shou M; Unger EF. Combined bromodeoxyuridine immunohistochemistry and Masson trichrome staining: facilitated detection of cell proliferation in viable vs. infarcted myocardium. Biotechnic and Histochemistry, 1992, 67(5):253–5. 27. Potten CS; Kellett M; Rew DA; Roberts SA. Proliferation in human gastrointestinal epithelium using bromodeoxyuridine in vivo: data for different sites, proximity to a tumour, and polyposis coli [see comments]. Gut, 1992, 33(4):524–9. 28. Potten CS; Kellett M; Roberts SA; Rew DA; Wilson GD. Measurement of in vivo proliferation in human colorectal mucosa using bromodeoxyuridine. Gut, 1992, 33(1):71–8. 29. Roncucci L; Pedroni M; Scalmati A; Bormioli ML; Sassatelli R; Fante R; Losi L; Di Gregorio C; Petocchi B; Ponz de Leon M. Cell kinetics evaluation of colorectal tumors after in vivo administration of bromodeoxyuridine. International Journal of Cancer, 1992 2, 52(6):856–61. 30. Shibuya M; Saito F; Miwa T; Davis RL; Wilson CB; Hoshino T. Histochemical study of pituitary adenomas with Ki-67 and anti-DNA polymerase alpha monoclonal antibodies, bromodeoxyuridine labeling, and nucleolar organizer region counts. Acta Neuropathologica, 1992, 84(2):178–83. 31. van de Kant HJ; de Rooij DG. Periodic acid incubation can replace hydrochloric acid hydrolysis and trypsin digestion in immunogold—silver staining of bromodeoxyuridine incorporation in plastic sections and allows the PAS reaction. Histochemical Journal, 1992, 24(3):170–5. 32. Yanik G; Yousuf N; Miller MA; Swerdlow SH; Lampkin B; Raza A. In vivo determination of cell cycle kinetics of non-Hodgkin’s lymphomas using iododeoxyuridine and bromodeoxyuridine. Journal of Histochemistry and Cytochemistry, 1992, 40(5):723–8. 33. Yu CC; Woods AL; Levison DA. The assessment of cellular proliferation by immunohistochemistry: a review of currently available methods and their applications. Histochemical Journal, 1992, 24(3):121–31. 34. Browman GP; Kanclerz A; Booker L; Daya D; Archibald SD; Young JE; Goldsmith CH. Optimal conditions for immunohistochemical determination of the in vitro DNA synthesis labelling index with bromodeoxyuridine in head and neck cancer. Cell Proliferation, 1991, 24(6):579–85. 35. Hirano T; Zitsch RP 3d; Gluckman JL. The use of bromodeoxyuridine cytokinetic studies as a prognostic indicator of cancer of the head and neck. Laryngoscope, 1991, 101(2):130–3. 36. Kitagawa K; Amagai M; Miyakawa S; Hayakawa K; Ueda M; Shimizu N. Trichilemmal tumor arising in a seborrheic keratosis: analysis of cell kinetics by BrdU staining. Journal of Dermatology, 1991, 18(6):334–9. 37. Kono T; Takada M; Wu JY; Kitai ST. Double immunohistochemical detection of transmitter phenotype of proliferating cells using bromodeoxyuridine. Neuroscience Letters, 1991, 132(1):113–6. 38. Labrecque C; Huard J; Dansereau G; Albert N; Tremblay JP. In vitro bromodeoxyuridine labeling of nuclei: application to myotube hybridization. Journal of Histochemistry and Cytochemistry, 1991, 39(10):1421–6. 39. Lacy ER; Kuwayama H; Cowart KS; King JS; Deutz AH; Sistrunk S. A rapid, accurate, immunohistochemical method to label proliferating cells in the digestive tract. A comparison with tritiated thymidine. Gastroenterology,1991, 100:259–62. 40. Lundy MW; Hendrix T; Wergedal JE; Baylink DJ. Growth factor-induced proliferation of osteoblasts measured by bromodeoxyuridine immunocytochemistry. Growth Factors, 1991, 4:257–64. 41. Migheli A; Mocellini C; Giordana MT. Ultrastructural detection of DNA-incorporated bromodeoxyuridine in resin embedded brain tissue. Histochemistry, 1991, 95(5):491–4. 42. Miller MA; Mazewski CM; Yousuf N; Sheikh Y; White LM; Yanik GA; Hyams DM; Lampkin BC; Raza A. Simultaneous immunohistochemical detection of IUdR and BrdU infused intravenously to cancer patients. Journal of Histochemistry and Cytochemistry, 1991, 39(4):407–12. 43. Morimura T; Kitz K; Stein H; Budka H. Determination of proliferative activities in human brain tumor specimens: a comparison of three methods. Journal of Neuro-Oncology, 1991, 10:1–11. 44. Smedman L; Joki A; Perlmann P; Troye-Blomberg M. Whole-blood microassay for immunodetection of antigen speci c cell mediated immunity using bromodeoxyuridine incorporation. Journal of Clinical and Laboratory Immunology, 1991, 34(2):79–85. 45. Soriano E; Del Rio JA; Martinez-Guijarro FJ; Ferrer I; Lopez C. Immunocytochemical detection of 5'-bromodeoxyuridine in uoro-gold-labeled neurons: a simple technique to combine retrograde axonal tracing and neurogenetic characterization of neurons. Journal of Histochemistry and Cytochemistry, 1991, 39(11):1565–70. 46. Tsujihashi H; Nakanishi A; Matsuda H; Uejima S; Kurita T. Cell proliferation of human bladder tumors determined by BrdUrd and Ki-67 immunostaining. J of Urology, 1991, 145:846–9. 47. van Dierendonck JH; Wijsman JH; Keijzer R; van de Velde CJ; Cornelisse CJ. Cell-cycle-related staining patterns of anti-proliferating cell nuclear antigen monoclonal antibodies. Comparison with BrdUrd labeling and Ki-67 staining. American Journal of Pathology, 1991, 138(5):1165–72. 48. Apte SS. Validation of bromodeoxyuridine immunohistochemistry for localization of S-phase cells in decalci ed tissues. A comparative study with tritiated thymidine autoradiography. Histochemical Journal, 1990, 22(8):401–8. 49. Apte SS; Puddle B. Bromodeoxyuridine (BrdUrd) immunohistochemistry in undecalci ed plastic-embedded tissue. Elimination of the DNA denaturation step. Histochemistry, 1990, 93:631–5. 50. Detta A; Hitchcock E. Rapid estimation of the proliferating index of brain tumours. Journal of Neuro-Oncology, 1990, 8(3):245–53. 51. Fukuda K; Iwasaka T; Hachisuga T; Sugimori H; Tsugitomi H; Mutoh F. Immunocytochemical detection of S-phase cells in normal and neoplastic cervical epithelium by anti-BrdU monoclonal antibody. Analytical and Quantitative Cytology and Histology, 1990, 12(2):135–8. 52. Hayashi H; Ooi A; Nakanishi I. Biological and clinicopathological signi cance of endocrine differentiation of gastric adenocarcinoma evaluated by double immunohistochemical labeling for chromogranin A and bromodeoxyuridine. Japanese Journal of Clinical Oncology, 1990, 20(4):339–48. 53. Hume WJ; Keat S. Immunohistological optimization of detection of bromodeoxyuridine-labeled cells in decalci ed tissue. Journal of Histochemistry and Cytochemistry, 1990, 38:509–13. 54. Jirikowski GF; Ramalho-Ortigao JF; Kesse KW; Bloom FE. In situ hybridization of semithin Epon sections with BrdU labelled oligonucleotide probes. Histochemistry, 1990, 94(2):187–90. 55. Latos-Bielenska A; Trautmann T; Vogel W. Visualization of DNA in pachytene by monoclonal antibodies against BrdU reveals synaptonemal complex-like structures. Cytogenetics and Cell Genetics, 1990, 54(1–2):24–8. 56. McDowell EM; Zhang XM; DeSanti AM. Localization of healing tracheal wounds using bromodeoxyuridine immunohistochemistry. Stain Technology, 1990, 65(1):25–9. 57. Powell BL; Gregory BW; Kute TE; Morgan TM; Lyerly ES; Capizzi RL. Bromodeoxyuridine incorporation into DNA of human leukemia cells is not concentration dependent. Cytometry, 1990, 11(3):438–41. 58. Sapino A; Macri L; Gugliotta P; Bussolati G. Evaluation of proliferating cell types in human and mouse mammary gland by a double immunostaining procedure. Acta Histochemica. Supplementband, 1990, 40:81–4. 59. Scheres HM; Schutte B; De Goij AF; Bosman FT. Estrogen and progesterone receptor expression and proliferation in breast cancer cells, studied with ow cytometry. Acta Histochemica. Supplementband, 1990, 40:71–6. 60. Van de Kant HJ; Van Pelt AM; Vergouwen RP; De Rooij DG. A rapid immunogold-silver staining for detection of bromodeoxyuridine in large numbers of plastic sections, using microwave irradiation. Histochemical Journal, 1990, 22(6–7):321–6. 61. Vogel W; Mehnert K; Pentz S. Demonstration of chromosome replication by BrdU antibody technique and electron microscopy. Human Genetics, 1990, 84(3):237–40. 62. Welberg JW; de Vries EG; Hardonk MJ; Mulder NH; Harms G; Grond J; Zwart N; Koudstaal J; de Ley L; Kleibeuker JH. Proliferation rate of colonic mucosa in normal subjects and patients with colonic neoplasms: a re ned immunohistochemical method. Journal of Clinical Pathology, 1990, 43(6):453–6. 63. Williams LS; Nordberg JA; Chau PC. Evaluation of immunocytochemical detection methods of incorporated bromodeoxyuridine in hybridomas. Cytometry, 1990, 11(4):490–7.