SM1667PS Bromodeoxyuridine / BrDU antibody

See related secondary antibodies

Search for all "Bromodeoxyuridine / BrDU"

0.1 mg / €260.00
Please visit the country specific website of Acris Antibodies or contact your local Distributor to buy this product.

Quick Overview

Rat anti Bromodeoxyuridine / BrDU BU1/75

SM1667PS

More Views

  • SM1667PS
  • SM1667PS

Product Description for Bromodeoxyuridine / BrDU

Rat anti Bromodeoxyuridine / BrDU BU1/75.
Presentation: Purified
Product is tested for Immunocytochemistry/Immunofluorescence, Flow Cytometry, Paraffin Sections.

Properties for Bromodeoxyuridine / BrDU

Product Category Primary Antibodies
Target Category
Quantity 0.1 mg
Presentation Purified
Applications F, ICC/IF, P
Clonality Monoclonal
Clone BU1/75
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 SM15P, SM15PX
Application Flow Cytometry: Use 20 µl of 1/25-1/200 diluted antibody to label 106 cells in 100 µl. Membrane permeabilization is required (See protocol below).
Immunohistochemistry on Formalin-Fixed, Paraffin-Embedded Sections: 1/25-1/200. 
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, 10 mM 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. Pre-treatment of tissues with proteinase K should be avoided.
Immunocytochemistry: 1/50-1/100.
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 0.5 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

As a Positive Control, BrdU labelled cells maybe obtained from Phoenix Flow Systems (http://www.phnxflow.com), catalogue number ACNC12.

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.
Product Citations
Purchased from Acris:
  1. Sonal, Sidhaye J, Phatak M, Banerjee S, Mulay A, Deshpande O, et al. Myosin Vb mediated plasma membrane homeostasis regulates peridermal cell size and maintains tissue homeostasis in the zebrafish epidermis. PLoS Genet. 2014 Sep 18;10(9):e1004614. doi: 10.1371/journal.pgen.1004614. eCollection 2014 Sep. PubMed PMID: 25233349. (Free PMC Article available, 7 images available)
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.
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
Purification:
Affinity Chromatography on Protein G
Buffer System:
PBS
Preservatives:
0.09% Sodium Azide
State:
Liquid purified IgG fraction
Purified
Specificity
Specificity:
This antibdoy reacts with BrdU in sigle stranded DNA, BrdU attached to a protein carrier or free BrdU.
The antibody detects nucleated cells in S-phase with have had BrdU incorporated into their DNA.
It also reacts with Chlorodeocyuridine but with reduced staining.
The antibody does not cross react with Thymidine.

Accessory Products

  • LinkedIn