Immunofluorescent analysis of BrdU (green) showing staining in the nucleus of Hela cells treated with 100nM BrdU (right) and untreated Hela cells (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a BrdU monoclonal antibody (Product # MA3-071) in 3% BSA-PBS at a dilution of 1:50 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Chemical|
|Published species reactivity||Rat, Mouse, Human, Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||5-iodouridine covalently coupled to ovalbumin.|
|Storage buffer||0.01M HEPES with 0.15M NaCl, 30% glycerol|
|Contains||0.01% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100|
|Immunofluorescence (IF)||1:1000 - 1:2500|
|Immunohistochemistry (IHC)||1:1000 - 1:2500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA3-071 detects 5-Bromo-2-deoxyuridine (BrdU) in many samples.
MA3-071 has been successfully used in immunofluorescence, immunohistochemistry, and flow cytometry applications.
The MA3-071 immunogen is 5-iodouridine covalently coupled to ovalbumin.
This antibody was orginally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.
Bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) is a synthetic nucleoside that is an analogue of thymidine. BrdU is commonly used in the detection of proliferating cells in living tissues. BrdU can be incorporated into the newly synthesized DNA of replicating cells (during the S phase of the cell cycle), substituting for thymidine during DNA replication. Antibodies specific for BrdU can then be used to detect the incorporated chemical, thus indicating cells that were actively replicating their DNA. Binding of the antibody requires denaturation of the DNA, usually by exposing the cells to acid or heat.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
The antibodies against 5-bromo-2'-deoxyuridine specifically recognize trifluridine incorporated into DNA.
MA3-071 was used in flow cytometry to analyze the specific recognition of trifluridine incorporated into DNA by antibodies against 5-bromo-2'-deoxyuridine
|Kitao H,Morodomi Y,Niimi S,Kiniwa M,Shigeno K,Matsuoka K,Kataoka Y,Iimori M,Tokunaga E,Saeki H,Oki E,Maehara Y||Scientific reports (6:null)||2016|
Cdc42 overexpression induces hyperbranching in the developing mammary gland by enhancing cell migration.
MA3-071 was used in immunohistochemistry to study the role of cell migration in the mechanism by which overexpression of Cdc42 promotes developing mammary gland hyperbranching
|Bray K,Gillette M,Young J,Loughran E,Hwang M,Sears JC,Vargo-Gogola T||Breast cancer research : BCR (15:null)||2014|
Atorvastatin and whisker stimulation synergistically enhance angiogenesis in the barrel cortex of rats following focal ischemia.
MA3-071 was used in immunohistochemistry to study the synergistic effects of atorvastatin and whisker stimulation on rat barrel cortex angiogenesis following ischemia
|Zhang Y,Huang S,Wang B,Sun B,Li W,Lu X,Ding X||Neuroscience letters (525:135)||2012|
Dual mechanism for the translation of subgenomic mRNA from Sindbis virus in infected and uninfected cells.
MA3-071 was used in immunocytochemistry to study the translation of sub-genomic DNA in Sindbis-infected and -uninfected BHK cells and the different mechanisms utilized
|Sanz MA,Castelló A,Ventoso I,Berlanga JJ,Carrasco L||PloS one (4:null)||2009|
|Not Applicable||Not Cited||
In vivo proliferation and cell cycle kinetics of long-term self-renewing hematopoietic stem cells.
MA3-071 was used in immunocytochemistry to study the proliferation and cell cycle kinetics of long-term self-renewing murine long-term hematopoietic stem cells.
|Cheshier SH,Morrison SJ,Liao X,Weissman IL||Proceedings of the National Academy of Sciences of the United States of America (96:3120)||1999|