11296736001
Roche
5-Bromo-2′-deoxy-uridine Labeling and Detection Kit I
sufficient for ≤100 tests, kit of 1 (5 components), suitable for immunofluorescence
Sinónimos:
5-BrdU, 5-Bromo-2-deoxyuridine
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About This Item
Código UNSPSC:
41116133
Productos recomendados
uso
sufficient for ≤100 tests
Nivel de calidad
envase
kit of 1 (5 components)
fabricante / nombre comercial
Roche
técnicas
immunofluorescence: suitable
temp. de almacenamiento
−20°C
Descripción general
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.
Especificidad
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.
Aplicación
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.
Envase
1 kit containing 5 components.
Nota de preparación
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).
Otras notas
For life science research only. Not for use in diagnostic procedures.
Solo componentes del kit
Referencia del producto
Descripción
- 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
Palabra de señalización
Danger
Frases de peligro
Consejos de prudencia
Clasificaciones de peligro
Aquatic Chronic 3 - Eye Irrit. 2 - Muta. 1B - Skin Irrit. 2 - Skin Sens. 1
Código de clase de almacenamiento
12 - Non Combustible Liquids
Clase de riesgo para el agua (WGK)
WGK 2
Punto de inflamabilidad (°F)
does not flash
Punto de inflamabilidad (°C)
does not flash
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