BrdU (Bromodeoxyuridine) Antibody Review

 

 

 

Introduction

  • a thymidine analog that incorporates DNA of dividing cells during the S-phase of the cell cycle. However, BrdU is not a marker of the S-phase of the cell cycle. As a thymidine analog, it is a marker of DNA synthesis. PMID: 17020783

  • an exogenously administered DNA precursor label and used as a proliferation marker PMID: 7912188, PMID: 9268038

BrdU Incorporation Techniques

  • BrdU can be administered to laboratory animals via IP injections, is readily incorporated into nuclei during the DNA synthetic phase of the cell cycle, and is detected with an anti-BrdU antibody. PMID: 14623936

  • BrdU was instilled intraperitoneally and the animals were painlessly sacrificed between 1 hour and 10 days after the injection. PMID: 9153107

  • both dose and exposure time to BrdU may influence the final results when cell proliferation is assessed, the variations obtained clearly depending on the technique used for the immunological detection of BrdU-positive S-phase cells. PMID: 7734409

  • Each patient received a local injection of 300 mg to 500 mg of BrdU into the cervix, at the time of endocervical laser conization, to label S-phase cells in neoplastic tissue. Labeled cells in normal and neoplastic tissues were detected in biopsy specimens or smears by indirect immunoperoxidase staining using the anti-BrdU MAb. PMID: 2350389

  • Specimens were incubated in vitro with BrdUrd and then fixed and paraffin embedded. Sections were immunohistochemically stained with antibodies to BrdUrd, proliferating cell nuclear antigen (PCNA), and Ki-67. PMID: 7855002

Comparison of BrdU with Other Proliferation Markers (Ki-67, PCNA)

  • Comparison of cell proliferation index in equine and caprine embryos using a modified BrdU incorporation assay. PMID: 15907994

  • both ISH for histone mRNA and IHC with Ki-67 (MIB-5) are preferable techniques for assessment of cell proliferation in rat paraffin-embedded renewing tissues compared to PCNA IHC. They can substitute for BrdU IHC when necessary. PMID: 14623936

  • The BrdU and cyclin A indices were significantly correlated with each other. In the more dysplastic cases, the cyclin A LI was quantitatively much larger than that for BrdU, suggesting that the protein was being overexpressed. It was concluded that as a tool to study the kinetic aspects of the cell cycle in dysplastic lesions cyclin A was limited by the fact that it is overexpressed. In minimally dysplastic lesions and normal epithelia, however, cyclin A may be a viable alternative to BrdU for the study of the S-phase. PMID: 11016684

  • PCNA has a lower variation between biopsies than BrdU, but higher variation between scorings. When used in a clinical or epidemiological setting, it is important to take multiple biopsies at multiple time points. PMID: 10942872

  • MIB-1 is closely associated with BrdU in clinicopathologic findings and is a more useful tool for evaluating cell proliferation than PCNA. PMID: 9110347

  • The labelling indices determined by histone NISH and BrdU incorporation were similar, whereas that of Ki67 IHC was four times greater. PMID: 7718451

  • proliferation rate in livers of 120 mice (60 males and 60 females) was analyzed by immunohistochemical detection of bromodeoxyuridine (BrdU) incorporation and proliferating cell nuclear antigen (PCNA) expression on ethanol-fixed/paraffin-embedded specimens. PMID: 7987078

  • Double label immunofluorescence against PCNA and BrdU clearly revealed several characteristics of DNA replication sites in synchronized CHO cells. PMID: 7735116

  • PCNA could replace the BrdU method for identifying the proliferating cells, and the major advantages of PCNA method is that it could be done without any pretreatment and avoid injection of the teratogenic agent for diagnostic purpose. PMID: 1363339

  • percentage of PCNA-positive cells was correlated with the BrdU labelling index and the histological malignancy of the brain tumours. The correlation coefficient was 0.84. This suggests that the immunohistochemical staining for PCNA in paraffin sections is a good alternative to the BrdU labelling index. PMID: 1357920

Expression

  • BrdU is a reliable standard and a more useful tool for the evaluation of proliferative activity of breast tumors. PMID: 8822114

  • a significant increase in BrdU-positive cells in the neonatal mouse hippocampus in the injured area compared to the non-injured area, most prominent in the dentate gyrus (DG). PMID: 15743533

  • Increases in BrdU-positive cells were detected in the hippocampal dentate gyrus at 5, 7, and 9 days after ischemia. PMID: 15129187

Function

  • BrdU application does not induce apoptosis or necrosis as revealed with the annexin V/PI assay. We concluded that continuous BrdU treatment did not affect cell viability, but essentially alters cell cycle progression in three out of four cell lines tested. PMID: 15030553

  • BrdU labeling in combination with histopathological observation is therefore a reliable approach to assessment of test compound effects in vivo. PMID: 9035078

  • Incorporation of BrdU into the nuclei during a period of 4 h was used to determine the proliferation of the embryonic cells in vitro. PMID: 9227916

Applications

 

Electron Microscopy (EM)

  • A simplified method of immunostaining of bromodeoxyuridine (BrdU), a thymidine analogue, for routinely paraformaldehyde or glutaraldehyde and osmium tetroxide-fixed and epon embedded specimens was established using regenerating rat liver. PMID: 8811633

  • Demonstration of chromosome replication by BrdU antibody technique and electron microscopy. PMID: 2303245

ELISA

  • An ELISA-based method for the quantification of incorporated BrdU as a measure of cell proliferation in vivo. PMID: 16780957

  • a simple, sensitive, nonradioactive, relatively rapid and relatively inexpensive protocol to measure DNA synthesis in cultured cells by a chemiluminescent bromodeoxyuridine (BrdU) enzyme-linked immunosorbent assay (ELISA). PMID: 12609534

  • BrdU ELISA is the most sensitive of the three proliferation assays used for the assessment of CD4 + T lymphocyte growth and is the preferred assay when small changes in cell growth are expected. PMID: 10089097

Flow Cytometry (FC)

  • Three cell preparation protocols for BrdU measurement were compared for their ability to maintain fluorescent surface staining and scatter parameters of in vivo BrdU-labeled cells by flow cytometry. PMID: 16538653

  • bromodeoxyuridine (BrdU) incorporation assay by flow cytometry was compared with 3H-TdR incorporation assay. PMID: 12014586

  • cell kinetics were examined by flow cytometry with BrdU-PI double staining. PMID: 1284799

  • Flow cytometric bromodeoxyuridine (BrdU)/DNA analysis using fresh solid tumors and its clinical significance. PMID: 1447808

  • Bivariate distributions of BrdU and DNA content are simultaneously obtained by flow cytometric two parameter analysis with a double staining method. PMID: 1280304

  • Studies on antitumor effects and mechanism of action of l-hexylcarbamoyl-5-fluorouracil by DNA/BrdU double staining using flow cytometry. PMID: 1382573

  • a method for the estimation of the proliferative activity of tumour cells obtained by fine needle sampling without aspiration (FNS), using simultaneously S-phase fractions (SPF) measured on DNA histograms and 5-bromodeoxyuridine (BrdU) labelling index (BLI) measured by flow cytometry. PMID: 1911191

  • BrdU-Hoechst flow cytometry: a unique tool for quantitative cell cycle analysis. PMID: 2448151

  • Peripheral blood lymphocyte cultures of a patient with clinical evidence of Fanconi's anemia (FA) were investigated by means of BrdU/Hoechst flow cytometry. PMID: 3592130

  • Differences in growth factor sensitivity between primary and transformed murine cell cultures revealed by BrdU/Hoechst flow cytometry. PMID: 3327716

  • An improved BrdU-Hoechst flow assay was applied to cell kinetic studies of human lymphocyte cultures during a 24-96 hr interval after PHA stimulation. PMID: 4055906

  • Cell cycle kinetics by BrdU-Hoechst flow cytometry: an alternative to the differential metaphase labelling technique. PMID: 2411405

  • Flow cytometric analysis of chromosomes and cells using a modified BrdU-Hoechst method. PMID: 6184345

Immunocytochemistry

  • Cell proliferation was assessed immunocytochemically using BrdU incorporation technique and documented on both ganglionic sections and microglia cultured cells at different experimental conditions and times after activation. PMID: 15270245

  • Cells in S-phase of cell cycle were labelled in vitro by incubation of fresh tissue fragments with 5-bromo 2-deoxyuridine (BrdU), a thymidine analogue. Nuclei of cells in active DNA synthesis were stained by an anti-BrdU monoclonal antibody (Mab). PMID: 8217353

  • A new procedure is described to generate single-stranded DNA by exonuclease III (Exo III) digestion for bromodeoxyuridine (BrdU) immunocytochemistry on tissue sections. PMID: 1452452

  • Immunocytochemical detection of S-phase cells in normal and neoplastic cervical epithelium by anti-BrdU monoclonal antibody. PMID: 2350389

Immunohistochemistry (IHC)

  • DNAse I pre-treatment markedly enhances detection of nuclear cyclin-dependent kinase inhibitor p57Kip2 and BrdU double immunostaining in embryonic rat brain. PMID: 17024454

  • BrdU immunohistochemistry for studying adult neurogenesis: Paradigms, pitfalls, limitations, and validation. PMID: 17020783

  • Evaluation of cell proliferation in rat tissues with BrdU, PCNA, Ki-67(MIB-5) immunohistochemistry and in situ hybridization for histone mRNA. PMID: 14623936

  • Fixation in methacarn gave the highest labeling index (16.4%), which was comparable to that observed in unfixed frozen sections (17.5%). Formalin fixation alone dramatically suppressed the labeling index (0.3%), which was only partially recovered using various antigen retrieval techniques (2.1-8.1%). Methacarn fixation is recommended for prospective studies in which BrdU detection is planned because of the quantitative recovery of epitope and the simplicity of the approach. PMID: 10681389

  • bromodeoxyuridine (BrdU) immunohistochemistry in undecalcified adult rat tibiae to study cell kinetics in various bone compartments: primary and secondary spongiosae, periosteum, and bone marrow. PMID: 9283606

  • Image analysis of bromodeoxyuridine (BrdU) staining for measurement of S-phase in rat and mouse liver. PMID: 8014477

  • volume and DNA synthesis of the neuroepithelium in induced myeloschisis in Long-Evans rats as shown by hematoxylin-eosin and BrdU/antiBrdU immunohistochemical staining patterns were examined at different stages of embryonal development. PMID: 8044815

  • possibility of detecting in situ proliferating myosatellite cells during postlarval muscle growth in the carp (Cyprinus carpio, L.) by means of BrdU and PCNA (Cyclin) immunohistochemistry has been evaluated on paraffin embedded sections. PMID: 7524812

  • Colonic crypt cell proliferation was measured using a monoclonal antibody to bromodeoxyuridine by an immunohistochemical technique. PMID: 1463991

  • Immunohistochemical observations of dividing cells in olfactory epithelium using anti-BrdU antibody. PMID: 1716113