SM1667B Bromodeoxyuridine / BrDU antibody

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0.1 mg / €440.00
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Rat anti Bromodeoxyuridine / BrDU BU1/75 (ICR1)

SM1667B

Product Description for Bromodeoxyuridine / BrDU

Rat anti Bromodeoxyuridine / BrDU BU1/75 (ICR1).
Presentation: Biotin
Product is tested for Flow Cytometry.

Properties for Bromodeoxyuridine / BrDU

Product Category Primary Antibodies
Target Category
Quantity 0.1 mg
Presentation Biotin
Applications F
Clonality Monoclonal
Clone BU1/75 (ICR1)
Host Rat
Isotype IgG2a
Shipping to Worldwide
PDF datasheet View Datasheet
Manufacturer Acris Antibodies GmbH
Material safety datasheet MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Isotype control SM15B
Application Flow Cytometry: Use 20 µl of neat Antibody to label 106 cells in 100 µl. 
See Protocol for more details.
Background 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 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 S-phase cells may 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.
Concentration 1.0 mg/ml
Protocols Flow Cytometry Analysis
Prepare the following solutions before proceeding:
Phosphate buffered saline (PBS)
2N HCl containing 0.5% Triton X-100
PBS containing 0.05% Tween-20 
PBS containing 1% BSA (PBS/BSA)
10 mg/ml Propidium iodide (PI)
0.1 M Na2B4O7, pH 8.5 

1. Add BrdU to the cell suspension in culture medium to a final concentration of 10 µmol/L and incubate for 30 min in a CO2 incubator at 37°C. 
2. Wash cells twice with PBS/BSA by centrifuging at 500g for 10 min, decant supernatant and resuspend in a miniumum volume of PBS. 
3. Add cells slowly into 5 ml of 70% ethanol at -20°C, mixing continuously (vortex preferred). Incubate on ice for 30 min. 
4. Centrifuge at 500g for 10 min, decant supernatant, and resuspend cell pellet. 
5. Add 2 ml of 2N HCl containing 0.5% Triton X-100 and incubate the cells for 30 min at RT (preferably on a rocking platform). 
6. Centrifuge at 500g for 10 min, decant supernatant and resuspend in 3 ml of 0.1 M Na2B4O7, pH 8.5. 
7. Centrifuge at 500g for 10 min, decant supernatant and resuspend the cells in PBS/BSA + 0.05% Tween-20. Adjust cell concentration to 1 x 107/ml. 
8. Aliquot 100 ul of cell suspension into required number of 12 x 75 mm tubes. 
9. Incubate the cells with the BrdU antibody at the recommended dilution for 30 min at RT. 
10. Add 2 ml of PBS/BSA and centrifuge the cells at 1000 rpm for 5 min. 
11. If a secondary antibody layer is required then decant the supernatant and incubate the cells with the secondary antibody for 30 min at RT. If no secondary antibody layer is required then proceed to step 13. 
12. Wash the cells by repeating step 10. 
13. Decant off the supernatant and add 1ml of PBS containing 10 µg/ml PI (dilute the 10 mg/ml solution of PI 1/1000 in a suitable volume of PBS) 
14. Analyse cells by Flow Cytometry following the manufacturers instructions. The PI should be read on the appropriate channel set to the Peak/Area and not log scale.
General Readings 1. Vanderlaan, M. & Thomas, C.B. (1985) Characterization of monoclonal antibodies to bromodeoxyuridine. Cytometry. 6: 501-505. 
2. Ghiringelli, F. et al. (2005) Tumor cells convert immature myeloid dendritic cells into TGF-beta-secreting cells inducing CD4+CD25+ regulatory T cell proliferation. J. Exp. Med. 202: 919-929. 
3. Dolbeare, F. (1995) Bromodeoxyuridine: a diagnostic tool in biology and medicine, Part I: Historical perspectives, histochemical methods and cell kinetics. Histochem. J. 27: 339-369. 
4. Das, G. et al. (2009) Cyclin D1 fine-tunes the neurogenic output of embryonic retinal progenitor cells. Neural Dev. 
4: 15. 5. Nakhai, H. et al. (2008) Conditional ablation of Notch signaling in pancreatic development. Development. 135: 2757-65. 
6. Ghai, K. et al. (2010) Notch signaling influences neuroprotective and proliferative properties of mature Müller glia. J Neurosci. 30: 3101-12. 
7. Amador-Arjona, A. et al. (2011) Primary cilia regulate proliferation of amplifying progenitors in adult hippocampus: implications for learning and memory. J Neurosci. 31: 9933-44. 
8. Bugler, B. et al. (2010) Unscheduled expression of CDC25B in S-phase leads to replicative stress and DNA damage. Mol Cancer. 9: 29. 
9. Gonzalo-Gobernado, R. et al (2009) Mobilization of neural stem cells and generation of new neurons in 6-OHDA-lesioned rats by intracerebroventricular infusion of liver growth factor. J Histochem Cytochem. 57: 491-502 
10. Xu, Q. et al. (2010) Sonic hedgehog signaling confers ventral telencephalic progenitors with distinct cortical interneuron fates. Neuron. 65: 328-40. 
11. Zhang, J. et al. (2010) A powerful transgenic tool for fate mapping and functional analysis of newly generated neurons. BMC Neurosci. 11: 158. 
12. Bonzo, J.A. et al. (2012) Suppression of Hepatocyte Proliferation by Hepatocyte Nuclear Factor 4a in Adult Mice. J Biol Chem. Jan 12. [Epub ahead of print] 
13. Knopf, F. et al. (2011) Bone regenerates via dedifferentiation of osteoblasts in the zebrafish fin. Dev Cell. 20: 713-24. 
14. Grotek, B. et al. (2013) Notch signaling coordinates cellular proliferation with differentiation during zebrafish fin regeneration. Development. 140: 1412-23.
15. Kim, T.H. et al. (2011) Genetic evidence that intestinal notch functions vary regionally and operate through a common mechanism of math1 repression. J Biol Chem. Jan 31. [Epub ahead of print] 
16. Kroehne, V. et al. (2011) Regeneration of the adult zebrafish brain from neurogenic radial glia-type progenitors. Development. 138: 4831-41. 
17. Liu, M.T. et al. (2009) 5-HT4 receptor-mediated neuroprotection and neurogenesis in the enteric nervous system of adult mice. J Neurosci. 29: 9683-99. 
18. Lundgren, O. et al. (2011) Intestinal epithelial stem/progenitor cells are controlled by mucosal afferent nerves. PLoS One. 6: e16295. 
19. Muja, N. et al. (2011) Neural precursors exhibit distinctly different patterns of cell migration upon transplantation during either the acute or chronic phase of EAE: A serial MR imaging study. Magn Reson Med. Feb 8. [Epub ahead of print] 
20. Puverel, S. et al. (2011) RanBPM is essential for mouse spermatogenesis and oogenesis. Development. 138: 2511-21. 
21. Sabo, J.K. et al. (2011) Remyelination is altered by bone morphogenic protein signaling in demyelinated lesions. J Neurosci. 31: 4504-10. 
22. Scheys, J.O. et al. (2011) Evidence of adrenal failure in aging dax1-deficient mice. Endocrinology. 152: 3430-9. 
23. Wang, W. et al. (2013) Extracellular signal-regulated kinase 5 (ERK5) mediates prolactin-stimulated adult neurogenesis in the subventricular zone and olfactory bulb. J Biol Chem. 288: 2623-31 
24. Zemans, R.L. et al. (2013) Role of ß-catenin-Regulated CCN Matricellular Proteins in Epithelial Repair After Inflammatory Lung Injury. Am J Physiol Lung Cell Mol Physiol. Jan 11. [Epub ahead of print] 
25. Zimmerman, K.M. et al. (2013) Diminished origin licensing capacity specifically sensitises tumour cells to replication stress. Mol Cancer Res. Jan 30. [Epub ahead of print] 
26. Maden, M. et al. (2013) Proliferation zones in the axolotl brain and regeneration of the telencephalon. Neural Dev. 8: 1. 
27. Perez-Ruiz A et al. {beta}-catenin promotes self-renewal of skeletal-muscle satellite cells. J Cell Sci : (2008).
28. Iulianella A, Sharma M, Durnin M, Vanden Heuvel GB, Trainor PA. Cux2 (Cutl2) integrates neural progenitor development with cell-cycle progression during spinal cord neurogenesis. Development. 2008 Feb;135(4):729-41. doi: 10.1242/dev.013276. PubMed PMID: 18223201.
Storage Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.
Shelf life: one year from despatch.
Format
Label:
Biotin
Purification:
Affinity Chromatography on Protein G
Buffer System:
PBS
Preservatives:
0.09% Sodium Azide
Stabilizers:
1% BSA
State:
Liquid purified IgG fraction from Tissue Culture Supernatant
Specificity
Specificity:
Reacts with BrdU in single stranded DNA, BrdU attached to a protein carrier or free BrdU. The antibody detects nucleated cells in S-phase which have had BrdU incorporated into their DNA. It also reacts with Chlorodeoxyuridine but with reduced staining. The antibody does not cross react with Thymidine.
Anti BrdU antibody cross reacts with Chlorodeoxyuridine (CldU) but does not cross react with Thymidine or Iododeoxyuridine (Aten et al. 1992). BrdU, IdU and CldU are analogs of Thymidine, they can incorporate into DNA during DNA synthesis replacing Thymidine. Antibody detection of incorporated BrdU in cellular DNA is extensively referenced as an accurate method to monitor cell proliferation in vivo and in vitro. In cell proliferation assays BrdU staining is coupled with the use of a dye that binds total DNA such as Propidium Iodide (PI). BrdU can be administered diluted in the culture medium or, in vivo via intraperitoneal injection, subcutaneous osmotic pump implants (Tesfaiqzi et al. 2004) or in drinking water (Moser et al. 2004).
The BrdU antibody clone BU1/75 (ICR1) has been used to detect CldU to study the speed of DNA replication fork (Bugler et al. 2010), in the detection of CldU label retaining stem cells (Kimoto et al. 2008) and label retaining neurons (Murata et al. 2011).

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