11296736001
Roche
5-Bromo-2′-deoxy-uridine Labeling and Detection Kit I
sufficient for ≤100 tests, kit of 1 (5 components), suitable for immunofluorescence
Synonym(s):
5-BrdU, 5-Bromo-2-deoxyuridine
Sign Into View Organizational & Contract Pricing
All Photos(1)
About This Item
UNSPSC Code:
41116133
Recommended Products
usage
sufficient for ≤100 tests
Quality Level
packaging
kit of 1 (5 components)
manufacturer/tradename
Roche
technique(s)
immunofluorescence: suitable
storage temp.
−20°C
General description
The kit is used for the detection of BrdU incorporated into cellular DNA using immunofluorescence microscopy. It is used for the detection of DNA synthesis by either in vitro labeling of cells or organ cultures, or by in vivo labeling, in which frozen or paraffin-embedded tissue sections must be prepared prior to fixation.
Normally, binding of the antibody is only achieved by denaturation of the DNA. This is usually obtained by exposing the cells to acid, base, or heat. These procedures result in destruction of cell integrity, including cell morphology and surface and cytoplasmatic markers.
The BrdU Labeling and Detection Kit I avoids these problems. The antibody preparation contains specific nucleases which allows access to BrdU after fixation in acidic ethanol. Therefore also simultaneous detection of other markers (double staining) is possible.
Normally, binding of the antibody is only achieved by denaturation of the DNA. This is usually obtained by exposing the cells to acid, base, or heat. These procedures result in destruction of cell integrity, including cell morphology and surface and cytoplasmatic markers.
The BrdU Labeling and Detection Kit I avoids these problems. The antibody preparation contains specific nucleases which allows access to BrdU after fixation in acidic ethanol. Therefore also simultaneous detection of other markers (double staining) is possible.
Specificity
Anti-BrdU monoclonal antibody specifically binds to 5-bromo-2′-deoxy-uridine, and shows cross-reactivity with 5-iodo-2′-deoxy-uridine (10%). Anti-BrdU shows no cross-reactivity with 5-fluoro-2′-deoxy-uridine or any endogenous cellular component, such as thymidine or uridine.
Application
Samples prelabeled with BrdU are fixed with ethanol, then incubated with a monoclonal antibody to BrdU. The antibody to BrdU supplied with the kit contains an optimized mixture of nucleases. These nucleases generate single-stranded DNA fragments, which allow binding of the antibody to BrdU without destruction of the cellular morphology. A fluorescein-labeled antibody to mouse immunoglobulin is added, then bound to the anti-BrdU antibody. Subsequently, the sample is evaluated using an immunofluorescence microscope.
BrdU Labeling and Detection Kit has been used for the detection of 5-bromo-2′-deoxy-uridine (BrdU) incorporated into cellular DNA.
- Safe: No radioisotopes are used
- Easy to perform: Follows a standard immunofluorescence protocol
- Sensitive: Denaturation of DNA with nucleases allows for highly sensitive detection of BrdU
- Flexible: Allows double-labeling protocols
BrdU Labeling and Detection Kit has been used for the detection of 5-bromo-2′-deoxy-uridine (BrdU) incorporated into cellular DNA.
Packaging
1 kit containing 5 components.
Preparation Note
Cell proliferation may be studied by monitoring the incorporation of a radioisotope, [3H]-thymidine, into cellular DNA, followed by autoradiography. Alternatively, 5-bromo-2′-deoxy-uridine (BrdU) may be used instead of thymidine. Cells that have incorporated BrdU into DNA are easily detected using a monoclonal antibody against BrdU and an enzyme- or fluorochrome-conjugated second antibody.
Working solution: BrdU labeling medium
Dilute BrdU labeling reagent 1:1000 with sterile cell culture medium (final concentration 10μM).
Note: For in vivo labeling undiluted BrdU labeling reagent (1 to 2ml/100 g body weight) is needed.
Prepare shortly before use.
Anti-BrdU working solution
Dilute anti-BrdU solution 1:10 with Incubation buffer.
Prepare shortly before use.
Anti-mouse-Ig-fluorescein stock solution
Dissolve anti-mouse-Ig-fluorescein solution in 1ml double-dist. water.
Anti-mouse-Ig-fluorescein working solution
Dilute anti-mouse Ig-fluorescein stock solution 1:10 with PBS. If an extended storage is desired, add BSA (bovine serum albumin), 10 mg/ml.
Prepare shortly before use.
Washing buffer
Dilute Washing buffer concentrate (10x) (bottle 2) 1:10 with double-dist. water.
Storage conditions (working solution): BrdU labeling medium
Store undiluted (1000x) medium in aliquots at -15 to -25°C.
Anti-BrdU working solution
Store undiluted antibody at -15 to -25°C.
Anti-mouse-Ig-fluorescein stock solution
Stable at 2 to 8°C
Washing buffer
Stable at 2 to 8°C
Sample material: Cell culture: adherent cells, suspension cells, organ, or explant cultures. Tissue sections (after in vivo labeling with BrdU).
Working solution: BrdU labeling medium
Dilute BrdU labeling reagent 1:1000 with sterile cell culture medium (final concentration 10μM).
Note: For in vivo labeling undiluted BrdU labeling reagent (1 to 2ml/100 g body weight) is needed.
Prepare shortly before use.
Anti-BrdU working solution
Dilute anti-BrdU solution 1:10 with Incubation buffer.
Prepare shortly before use.
Anti-mouse-Ig-fluorescein stock solution
Dissolve anti-mouse-Ig-fluorescein solution in 1ml double-dist. water.
Anti-mouse-Ig-fluorescein working solution
Dilute anti-mouse Ig-fluorescein stock solution 1:10 with PBS. If an extended storage is desired, add BSA (bovine serum albumin), 10 mg/ml.
Prepare shortly before use.
Washing buffer
Dilute Washing buffer concentrate (10x) (bottle 2) 1:10 with double-dist. water.
Storage conditions (working solution): BrdU labeling medium
Store undiluted (1000x) medium in aliquots at -15 to -25°C.
Anti-BrdU working solution
Store undiluted antibody at -15 to -25°C.
Anti-mouse-Ig-fluorescein stock solution
Stable at 2 to 8°C
Washing buffer
Stable at 2 to 8°C
Sample material: Cell culture: adherent cells, suspension cells, organ, or explant cultures. Tissue sections (after in vivo labeling with BrdU).
Other Notes
For life science research only. Not for use in diagnostic procedures.
Kit Components Only
Product No.
Description
- BrdU Labeling Reagent, sterile 1,000x concentrated
- Washing Buffer concentrate 10x concentrated
- Incubation Buffer
- Anti-BrdU antibody, contains nucleases for DNA denaturation
- Anti-mouse-Ig-fluorescein antibody
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Aquatic Chronic 3 - Eye Irrit. 2 - Muta. 1B - Skin Irrit. 2 - Skin Sens. 1
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 2
Flash Point(F)
does not flash
Flash Point(C)
does not flash
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
Already Own This Product?
Find documentation for the products that you have recently purchased in the Document Library.
Customers Also Viewed
Sandra Bauer et al.
The Journal of clinical endocrinology and metabolism, 89(2), 812-822 (2004-02-07)
We have tested the hypothesis that elevated concentrations of TNF alpha could impair trophoblast invasion. Using first-trimester placental explant cultures, we have demonstrated that the cytokine inhibits in vitro migration of extravillous trophoblasts (EVT) on collagen I, and invasion through
Boaz Kigel et al.
Blood, 118(15), 4285-4296 (2011-08-13)
Plexin-A4 is a receptor for sema6A and sema6B and associates with neuropilins to transduce signals of class-3 semaphorins. We observed that plexin-A1 and plexin-A4 are required simultaneously for transduction of inhibitory sema3A signals and that they form complexes. Unexpectedly, inhibition
Nitin Sabherwal et al.
Development (Cambridge, England), 136(16), 2767-2777 (2009-07-28)
During neurogenesis in Xenopus, apicobasally polarised superficial and non-polar deep cells take up different fates: deep cells become primary neurons while superficial cells stay as progenitors. It is not known whether the proteins that affect cell polarity also affect cell
Christoph Mück et al.
The journals of gerontology. Series A, Biological sciences and medical sciences, 65(11), 1165-1180 (2010-08-03)
Tumor necrosis factor-like cytokine 1A (TL1A) is expressed in endothelial cells and contributes to T-cell activation, via an extracellular fragment TL1A(L72-L251), generated by ectodomain shedding. Fragments of TL1A, referred to as vascular endothelial growth inhibitor, were found to induce growth
Bin Lin et al.
Frontiers in endocrinology, 8, 253-253 (2017-10-27)
With recent advances in stem cell technology, it is becoming efficient to differentiate human pluripotent stem cells (hPSCs) into cardiomyocytes, which can subsequently be used for myriad purposes, ranging from interrogating mechanisms of cardiovascular disease, developing novel cellular therapeutic approaches
Articles
Cell based assays for cell proliferation (BrdU, MTT, WST1), cell viability and cytotoxicity experiments for applications in cancer, neuroscience and stem cell research.
Cell based assays for cell proliferation (BrdU, MTT, WST1), cell viability and cytotoxicity experiments for applications in cancer, neuroscience and stem cell research.
Cell based assays for cell proliferation (BrdU, MTT, WST1), cell viability and cytotoxicity experiments for applications in cancer, neuroscience and stem cell research.
Cell based assays for cell proliferation (BrdU, MTT, WST1), cell viability and cytotoxicity experiments for applications in cancer, neuroscience and stem cell research.
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
Contact Technical Service