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Anti-BrdU Antibody, clone 131-14871

产品编号:4090842
规格:clone 131-14871, Chemicon®, from mouse
包装规格:100 μG
产品类别:进口试剂
品牌:Sigma-Aldrich
优惠价:立即咨询
产品价格
产品编号包装单位单价(元)国内现货国外库存询价单
4090842100 μG5520
产品别名

Anti-BrdU Antibody, clone 131-14871

brdu

基本信息
eCl@ss
32160702
NACRES
NA.41
General description【一般描述】
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.
Specificity【特异性】
Recognizes BrdU (Bromodeoxyuridine).
Application【应用】
Research Sub Category
Cell Cycle, DNA Replication & Repair
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.
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.
Research Category
Epigenetics & Nuclear Function
Physical form【外形】
Format: Purified
Protein A purified
Liquid in 10 mM Phosphate buffer, pH 7.4 containing 150 mM NaCl and 0.1% sodium azide.
Analysis Note【分析说明】
Control
Brdu treated cells
Other Notes【其他说明】
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information【法律信息】
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
产品性质
Quality Level【质量水平】
100
biological source【生物来源】
mouse
antibody form【抗体形式】
purified immunoglobulin
antibody product type
primary antibodies
clone【克隆】
131-14871, monoclonal
species reactivity (predicted by homology)
all
manufacturer/tradename
Chemicon®
technique(s)
flow cytometry: suitable
immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
isotype【同位素/亚型】
IgG1
shipped in【运输】
wet ice
产品说明
Storage and Stability【储存及稳定性】
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.
Disclaimer【免责声明】
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.
安全信息
Storage Class Code【储存分类代码】
12 - Non Combustible Liquids
WGK
WGK 2
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