MAB4072
Anti-BrdU Antibody, clone 131-14871
clone 131-14871, Chemicon®, from mouse
Synonym(e):
BrdU
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About This Item
UNSPSC-Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Empfohlene Produkte
Biologische Quelle
mouse
Qualitätsniveau
Antikörperform
purified immunoglobulin
Antikörper-Produkttyp
primary antibodies
Klon
131-14871, monoclonal
Speziesreaktivität (Voraussage durch Homologie)
all
Hersteller/Markenname
Chemicon®
Methode(n)
flow cytometry: suitable
immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
Isotyp
IgG1
Versandbedingung
wet ice
Posttranslationale Modifikation Target
unmodified
Allgemeine Beschreibung
The thymidine analog 5-bromo-2′-deoxyuridine (BrdU) is a common reagent used for both cell proliferation assays and for the detection of apoptotic cells. BrdU is readily incorporated into newly synthesized DNA, labeling cells that have progressed through the S-phase of the cell cycle and thereby serving as a marker for proliferating cells.
Spezifität
Recognizes BrdU (Bromodeoxyuridine).
Anwendung
Research Category
Epigenetik & nukleäre Funktionen
Epigenetik & nukleäre Funktionen
Research Sub Category
Zellzyklus, DNA-Replikation & -Reparatur
Zellzyklus, DNA-Replikation & -Reparatur
Anti-BrdU Antibody, clone 131-14871 is a Mouse Monoclonal Antibody for detection of Bromodeoxyuridine also known as BrdU & has been validated in FC, IHC, IHC(P). This bromodeoxyuridine antibody is expect to react in all species.
Immunohistochemistry on deparaffinized sections of kidney, duodenum, and liver tissue: 1:1000-1:1250.
Flow cytometry on 100μL of 106 cells: 1:5000.
Optimal dilutions must be determined by the end user.
IMMUNOHISTOCHEMICAL PROCEDURE
FOR PARAFFIN EMBEDDED TISSUE SECTIONS
STAINING PROCEDURE:
1. Deparaffinize sections in xylene 3 x 5 minutes. Resin embedded or GMA (Glycol methacrylate, 2-hydroxyethyl methacrylate) treated sections it is unnecessary to deparaffinize as there is no paraffin.
2. Place slides in graded alcohols (100, 95, 90%) to water at 2 minute intervals, and air dry. (Omit for GMA sections)
3. Circle sections with PAP pen or diamond pen to identify and dry thoroughly.
4. For GMA or resin sections, place slides in 0.5% Tween/PBS (pH 7.6) for 29 minutes.
5. Incubate in prewarmed (40°C) 1N HCl for 1 hour.
6. Wash 3 x 3 minutes with PBS (pH 7.6). Slides may be held at this point and the procedure continued the following day.
7. Incubate in 1X Trypsin solution (0.02-0.05% w/v, prewarmed to 40°C) for 20 minutes at room temperature for NBF (normal buffered formalin) fixed tissue or 10 minutes for Carnoy′s fixed tissue.
8. Wash 3 x 5 minutes with PBS (pH 7.6).
9. Incubate sections in 1% H2O2 (10 mL 30% H2O2 in 290 mL methanol) for 20 minutes, or GMA sections in 3% H2O2 (10 mL 30% H2O2 in 90 mL water) for 3 minutes.
10. Wash 2 x 2 minutes with PBS (pH 7.6).
11. Using the Vector ABC kit, add 3 drops of normal horse serum to 10 mL of PBS. Use this solution to block the slides for 20 minutes at room temperature.
12. Blot the slides of excess normal horse serum and incubate with MAB4072 diluted in PBS/BSA/Tween 20 for 1 hour at room temperature.
13. Wash 3 x 3 minutes with PBS (pH 7.6).
14. Add 3 drops of normal horse serum to 10 mL of PBS and then add 1 drop of biotinylated antibody stock. Incubate the slides with this diluted second antibody for 30 minutes at room temperature.
15. Prepare the Vectastain ABC Reagent by adding 2 drops of reagent A to 5 mL of PBS. Then add 2 drops of Reagent B to the same mixing bottle. Allow to sit for about 30 minutes prior to use.
16. Wash 3 x 3 minutes with PBS (pH 7.6).
17. Incubate the slides with ABC Reagent for 30 minutes at room temperature.
18. Wash 3 x 3 minutes with PBS (pH 7.6).
19. Prepare the substrate by mixing an equal volume of 0.02% H2O2 (made in distilled water from a 30% stock solution) and 0.1% DAB made in 0.1 M Tris buffer, pH 7.2. The H2O2 should be freshly prepared from concentrated stock. Because many peroxide substrates are unstable in the presence of H2O2 or when exposed to light, the substrate should be prepared just prior to use. Since DAB is a suspected carcinogen, care should be taken in handling and disposing of all peroxidase substrates.
20. Stain the slides with DAB substrate for 2-5 minutes.
21. Wash 2 x 2 minutes with distilled water.
22. Counterstain with hemotoxylin, dehydrate through xylene, and mount the coverslip.
Flow cytometry on 100μL of 106 cells: 1:5000.
Optimal dilutions must be determined by the end user.
IMMUNOHISTOCHEMICAL PROCEDURE
FOR PARAFFIN EMBEDDED TISSUE SECTIONS
STAINING PROCEDURE:
1. Deparaffinize sections in xylene 3 x 5 minutes. Resin embedded or GMA (Glycol methacrylate, 2-hydroxyethyl methacrylate) treated sections it is unnecessary to deparaffinize as there is no paraffin.
2. Place slides in graded alcohols (100, 95, 90%) to water at 2 minute intervals, and air dry. (Omit for GMA sections)
3. Circle sections with PAP pen or diamond pen to identify and dry thoroughly.
4. For GMA or resin sections, place slides in 0.5% Tween/PBS (pH 7.6) for 29 minutes.
5. Incubate in prewarmed (40°C) 1N HCl for 1 hour.
6. Wash 3 x 3 minutes with PBS (pH 7.6). Slides may be held at this point and the procedure continued the following day.
7. Incubate in 1X Trypsin solution (0.02-0.05% w/v, prewarmed to 40°C) for 20 minutes at room temperature for NBF (normal buffered formalin) fixed tissue or 10 minutes for Carnoy′s fixed tissue.
8. Wash 3 x 5 minutes with PBS (pH 7.6).
9. Incubate sections in 1% H2O2 (10 mL 30% H2O2 in 290 mL methanol) for 20 minutes, or GMA sections in 3% H2O2 (10 mL 30% H2O2 in 90 mL water) for 3 minutes.
10. Wash 2 x 2 minutes with PBS (pH 7.6).
11. Using the Vector ABC kit, add 3 drops of normal horse serum to 10 mL of PBS. Use this solution to block the slides for 20 minutes at room temperature.
12. Blot the slides of excess normal horse serum and incubate with MAB4072 diluted in PBS/BSA/Tween 20 for 1 hour at room temperature.
13. Wash 3 x 3 minutes with PBS (pH 7.6).
14. Add 3 drops of normal horse serum to 10 mL of PBS and then add 1 drop of biotinylated antibody stock. Incubate the slides with this diluted second antibody for 30 minutes at room temperature.
15. Prepare the Vectastain ABC Reagent by adding 2 drops of reagent A to 5 mL of PBS. Then add 2 drops of Reagent B to the same mixing bottle. Allow to sit for about 30 minutes prior to use.
16. Wash 3 x 3 minutes with PBS (pH 7.6).
17. Incubate the slides with ABC Reagent for 30 minutes at room temperature.
18. Wash 3 x 3 minutes with PBS (pH 7.6).
19. Prepare the substrate by mixing an equal volume of 0.02% H2O2 (made in distilled water from a 30% stock solution) and 0.1% DAB made in 0.1 M Tris buffer, pH 7.2. The H2O2 should be freshly prepared from concentrated stock. Because many peroxide substrates are unstable in the presence of H2O2 or when exposed to light, the substrate should be prepared just prior to use. Since DAB is a suspected carcinogen, care should be taken in handling and disposing of all peroxidase substrates.
20. Stain the slides with DAB substrate for 2-5 minutes.
21. Wash 2 x 2 minutes with distilled water.
22. Counterstain with hemotoxylin, dehydrate through xylene, and mount the coverslip.
Physikalische Form
Protein A purified
Format: Purified
Liquid in 10 mM Phosphate buffer, pH 7.4 containing 150 mM NaCl and 0.1% sodium azide.
Lagerung und Haltbarkeit
Maintain for 2 years at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Hinweis zur Analyse
Control
Brdu treated cells
Brdu treated cells
Sonstige Hinweise
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Rechtliche Hinweise
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Haftungsausschluss
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Lagerklassenschlüssel
12 - Non Combustible Liquids
WGK
WGK 2
Flammpunkt (°F)
Not applicable
Flammpunkt (°C)
Not applicable
Analysenzertifikate (COA)
Suchen Sie nach Analysenzertifikate (COA), indem Sie die Lot-/Chargennummer des Produkts eingeben. Lot- und Chargennummern sind auf dem Produktetikett hinter den Wörtern ‘Lot’ oder ‘Batch’ (Lot oder Charge) zu finden.
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