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5-Bromo-2-deoxy-uridine Labeling and Detection Kit I 11296736001
General descriptionThe 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.
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.ApplicationSamples 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.- 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).Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, usage sufficient for ≤100 tests Торговая марка Roche shipped in dry ice storage temp. −20°C
Safety Informationpictograms GHS07 signalword Warning hcodes H315 - H317 - H319 - H412 pcodes P261 - P264 - P273 - P280 - P333 + P313 - P337 + P313 RIDADR NONH for all modes of transport WGK Germany WGK 2 Flash Point F does not flash Flash Point C does not flash Узнать цену -
5-Bromo-2-deoxy-uridine Labeling and Detection Kit II 11299964001
General descriptionImmunohistocytochemical assay for the detection of 5-bromo-2′-deoxy-uridine (BrdU) incorporated into cellular DNA.
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.
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.ApplicationThe kit is used for the detection of BrdU incorporated into cellular DNA by immunohistocytochemistry.
5-Bromo-2-deoxy-uridine Labeling and Detection Kit II has been used in:- labeling of tooth roots for histology
- immunostaining of mice frontal sections
- immunofluorescence imaging of hepatocellular carcinoma sections
Features and Benefits- Safe: No radioisotopes are used.
- Easy to perform: Follows a standard immunohistochemistry protocol.
- Sensitive: Denaturation of DNA with nucleases allows for highly sensitive detection of BrdU.
- Flexible: Allows double-labeling protocols.
Packaging
1 kit containing 7 components.
Principle
Samples prelabeled with BrdU are fixed with ethanol, then incubated with a monoclonal antibody to BrdU, which contains an optimized mixture of nucleases. These nucleases generate single-stranded DNA fragments that allow binding of the antibody to BrdU. Next, an alkaline phosphatase (AP)-labeled antibody to mouse immunoglobulin is added, then bound to the anti-BrdU antibody. The sample is then incubated with the AP substrate and NBT/ BCIP, which is metabolized to form a colored reaction product. The sample is evaluated using a phase-contrast microscope.
Preparation Note
Working concentration: Working concentration of the labeling reagent corresponds to the WC of the In Situ Cell Proliferation Kits.
Sample material:
Cell culture: adherent cells, suspension cells, organ or explant cultures. Frozen or paraffin-embedded tissue sections (after in vivo labeling).Other NotesFor life science research only. Not for use in diagnostic procedures.ПараметрыQuality Level 100, usage sufficient for ≤100 tests Торговая марка Roche shipped in dry ice storage temp. −20°C
Safety Informationpictograms GHS02,GHS07,GHS08 signalword Danger hcodes H226 - H312 + H332 - H317 - H319 - H360D pcodes P201 - P210 - P261 - P280 - P308 + P313 - P370 + P378 WGK Germany WGK 2 Flash Point F 136.4 °F Flash Point C 58 °C Узнать цену -
5-Bromo-2-deoxy-uridine Labeling and Detection Kit III 11444611001
General descriptionProliferation in cell populations may be studied by incorporating the radioisotope [3H]-thymidine into cellular DNA. The amount of radioactive thymidine incorporated is determined by scintillation counting. Alternatively , 5-bromo-2-deoxy-uridine may be incorporated into cellular DNA. The amount of BrdU incorporated is determined by a standard ELISA protocol, which involves "tagging" the incorporated nucleotide with an anti-BrdU antibody. 96-well microplate cell ELISA for the detection of 5-bromo-2-deoxy-uridine (BrdU) incorporated into cellular DNA. Nonradioactive alternative for [3H]-thymidine based DNA synthesis and cell proliferation assays.
Contents- BrdU Labeling Reagent, 1,000x concentrated
- Washing Buffer concentrate, 10x concentrated
- Incubation Buffer
- Nucleases
- Anti-BrdU-POD, Fab fragments
- Substrate Buffer
- ABTS Substrate
- Substrate Enhancer
Specificity
Anti-BrdU-peroxidase, Fab fragments, specifically bind to 5-bromo-2′-deoxy-uridine incorporated into DNA. It shows no cross-reactivity with any endogenous cellular components, such as thymidine or uridine.ApplicationThe kit is used for the quantitative determination of BrdU incorporated into cellular DNA using a 96-well microplate cell ELISA format.
5-Bromo-2-deoxy-uridine Labeling and Detection Kit III has been used in cell proliferation assay, BrdU assay and proliferation assay.
Features and Benefits- Safer, since the kit does not use radioisotopes.
- Accurate, since results generated with this assay strongly correlate to those obtained with the [3 H]-thymidine method. (See Figure 2 below.)
- Sensitive. This assay and the [3 H]-thymidine assay are equally sensitive. (See Figure 2 below.)
- Fast, since results can be read with a multi-well ELISA reader, allowing simultaneous processsing of a large number of samples
- Easy, since the assay uses a standard cell ELISA protocol.
- Economical, since the assay requires no expensive equipment or additional reagents (e.g ., scintillation fluid).
Packaging
1 kit containing 8 components.
Principle
Cells cultured in a 96-well microplate are incubated with BrdU (see Performance). The labeled cells are fixed with ethanol. Prior to incubation with a monoclonal antibody to BrdU, DNA is partially digested with nucleases to allow the antibody to access BrdU. Next, the anti-BrdU antibody [labeled with peroxidase (POD)] is added. Finally, the POD substrate ABTS is added. POD catalyzes the cleavage of ABTS, producing a colored reaction product. The absorbance of the samples (at approximately 405 nm) is determined with a standard microplate (ELISA) reader.
Preparation Note
Working solution: BrdU labeling solution
Dilute BrdU labeling reagent 1 : 90 with sterile PBS or culture medium (resulting concentration: 111 µM BrdU) [e.g., for one 96-well microplate containing 100 µl medium per well, dilute 12 µl BrdU labeling reagent with 1.068 ml sterile PBS].
Note: The BrdU labeling solution should be prepared freshly before use.
Washing buffer
Dilute washing buffer concentrate (10x) 1 : 10 with double-dist. water [e.g., for one 96-well microplate dilute 9 ml washing buffer concentrate (10x) with 81 ml double-dist. water].
Note: If precipitates in Washing buffer, 10x conc. are visible, please incubate the bottle for 10 minutes at 37 °C in a water bath before you prepare Solution II.
Washing buffer is used to:- Prepare the anti-BrdU-peroxidase, working solution
- Wash cells after incubation with anti-BrdU-peroxidase
Incubation buffer- Ready-to-use
- Used to dilute the nucleases
Nucleases, stock solution
Reconstitute the nucleases in 1.3 ml double-dist. water containing 50% glycerol (w/v).
Nucleases, working solution
Dilute nucleases, stock solution, 1 : 100 with incubation buffer (e.g., for one 96-well microplate dilute 100 µl nucleases, stock solution, with 9.9 ml incubation buffer).
Anti-BrdU-peroxidase, Fab fragments, stock solution
Dissolve anti-BrdU-peroxidase, Fab fragments in 1.25 ml double-dist. water (final concentration: 20 U/ml).
Anti-BrdU-peroxidase, Fab fragments, working solution
Prepare anti-BrdU-peroxidase, Fab fragments, working solution shortly before use. Dilute anti-BrdU-peroxidase, Fab fragments, stock solution 1 : 100 with washing buffer supplemented with 10 mg/ml BSA (bovine serum albumin), [e.g., for one 96-well microplate dilute 100 µl anti-BrdU-peroxidase, Fab fragments, stock solution, with 9.9 ml PBS and BSA (final concentration: 200 mU/ml)].
Peroxidase substrate
Dissolve the ABTS powder in substrate buffer and stir at 15 to 25 °C to obtain a clear solution.
Peroxidase substrate containing substrate enhancer
If a low signal is expected, take an appropriate aliquot of substrate solution and add substrate enhancer , 1 mg/ml and dissolve by stirring for 15 minutes at 15 to 25 °C (e.g., for one 96-well microplate dissolve 10 mg substrate enhancer in 10 ml peroxidase substrate)
Note: The substrate solution containing substrate enhancer is stable for only 4 hours and should, therefore, be freshly prepared before use.
Storage conditions (working solution): BrdU labeling reagent
The undiluted BrdU labeling reagent (1000x) is stable at 2 to 8 °C for 6 months. It is stable stored in aliquots at -15 to -25 °C.
Washing buffer
Stable at 2 to 8 °C for 3 months.
Incubation buffer
Stable at 2 to 8 °C until the expiration date printed on the label
Nucleases, stock solution
Stable at -15 to -25 °C for 6 months.
Nucleases, working solution
must be prepared shortly before use.
Anti-BrdU-Peroxidase, Fab fragments, stock solution
Stable at 2 to 8 °C for 6 months. For long term storage it is recommended to store the solution in aliquots at -15 to -25 °C.
Anti-BrdU-Peroxidase, Fab fragments, working solution
must be prepared freshly before use.
Peroxidase substrate
Stable at 2 to 8 °C for 2 months when stored protected from light.
Peroxidase substrate containing substrate enhancer
must be prepared freshly before useOther NotesFor life science research only. Not for use in diagnostic procedures.Параметрыusage sufficient for ≤1,000 tests Quality Level 100 specific activity 10000 (Nuclease activity (vial 4)) Торговая марка Roche shipped in wet ice storage temp. 2-8°C
Safety Information
