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merck millipore,默克密理博,MAB3424,Anti-BrdU Antibody, clone AH4H7-1 / 131-14871
产品名称：Anti-BrdU Antibody, clone AH4H7-1 / 131-14871
产品型号：MAB3424
Use Anti-BrdU Antibody, clone AH4H7-1 / 131-14871 (Mouse Monoclonal Antibody) validated in FC, ICC, IHC to detect Bromodeoxyuridine also known as BrdU.
merck millipore,默克密理博,MAB3424,Anti-BrdU Antibody, clone AH4H7-1 / 131-14871
- 产品介绍
重要规格表
品种反应性 主要应用 宿主 格式 抗体类型 A IHC, FC, ICC M Purified Monoclonal Antibody 描述 产品目录编号 MAB3424 Replaces MAB1467 品牌系列 Chemicon®
商名 - Chemicon
描述 Anti-BrdU Antibody, clone AH4H7-1 / 131-14871 Alternate Names - BrdU
背景信息 Bromodeoxyuridine (BrdU) is a thymidine analog and is specifically incor-porated into DNA during DNA synthesis. Anti-bromodeoxyuridine monoclonal antibody is used to identify cells that have incorporated BrdU. This immunological detection scheme has several advantages over the use of radioactive thymidine incorporation for identifying cells under-going replication. Labeling and detection can be performed the same day instead of waiting several days, as required for autoradiography of tritium-labeled cells, and the necessity of using multiple specimens for obtaining the optimal exposure time is eliminated. In addition, anti-bromodeoxyuridine staining with flow cytometric analysis allows multiple parameters to be evaluated simultaneously. Anti-bromodeoxyuridine monoclonal antibody has been used for identi-fying proliferating cells in blood (Campana et al., 1988), tissues (Schutte et al., 1987; Hayashi, et al., 1988), tumors (Hoshino et al., 1986; Morstyn et al., 1983), as well as for determining plasma cell labeling indices (Greipp et al., 1985). 产品信息 格式 Purified 控制 - After incorporation of BrdU, all DNA containing species
演示 Liquid in 10 mM Phosphate buffer, pH 7.4 containing 150 mM NaCl and 0.1% sodium azide 应用 应用 Use Anti-BrdU Antibody, clone AH4H7-1 / 131-14871 (Mouse Monoclonal Antibody) validated in FC, ICC, IHC to detect Bromodeoxyuridine also known as BrdU. 主要应用 - Immunohistochemistry
- Flow Cytometry
- Immunocytochemistry
应用说明 Immunohistochemistry: (6 μg/ml)
Flow Cytometry: (0.2 μg/100 μl/10E6 cells) Optimal working dilutions must be determined by end user.
APPLICATIONS
Flow cytometry:The method below is based on that of M. Vanderlaan et al. (1986). Variations of this method exist in the literature, one consideration being the effect various fixation procedures have on the light-scattering properties of different cell populations. Procedure:
1. To label cells, pulse with 10 μM bromodeoxyuridine for 30 minutes. Harvest cells from culture.
2. Fix cells in 70% ethanol at +2-8°C for at least 30 min. Extract histones by resuspending cells in 1 mL chilled 0.1 M HCI containing 0.5% Triton X-100; incubate the suspension on ice for 10 minutes. Dilute acid with 5 mL distilled water and centrifuge at 200 x g for 10 min. Resuspend cells in 2 mL distilled water.
3. Denature cellular DNA by submerging the cell suspension into a boiling water bath for 10 min. Afterwards, quickly cool by placing the cell suspension in an ice slurry for several minutes. Wash cells in PBS that contains 0.5% Triton X-100.
4. Resuspend the cells (1-2 x 10 6 cells) in 100 μL of solution containing approximately 2 μg/mL anti-bromodeoxyuridine antibody diluted in PBS containing 0.1% BSA (0.2 μg/test). Incubate for 30 min at room temperature. Wash cells with PBS.
5. Resuspend cells in 100 μL of diluted goat anti-mouse IgG-FlTC Wash cells with PBS.
APPLICATIONS (Cont.)
Immunohistochemistry: Below is a procedure for staining cells that have been labeled with BrdU in vivo or in vitro. The procedure is based on the methods of B. Schutte et al. (1987) and D. Campana et al. (1988).
Preparation of tissue:
Inject animal with 50 mg BrdU/kg body weight. Sacrifice animal one hour later and remove organ or tissue under study. Embed tissue in OCT medium and snap-freeze by immersion into liquid nitrogen.Cut 4 mm frozen sections with a cryostat. Place sections on either albumin- or gelatin-coated slides.
Preparation of cells:
Pulse cells with 10 mM BrdU for 60 min. Cells grown on coverslips, or cytocentrifuge preparations made from cells grown in suspension, can be used for anti-bromodeoxyuridine staining according to the procedure below.
Procedure
1. Fix tissue sections or cells (on slide or coverglass) by immersing in absolute methanol for 10 minutes at +2-8°C. Air dry after removing from fixative. The slides can be stored at -20°C in a sealed box, or rehydrated to prepare for the assay procedure. To rehydrate, immerse in PBS for 3 min.
2. Denature DNA by incubating the slides in 2 N HCI for 60 min at +37°C.
3. Neutralize the acid by immersing the slides in 0.1 M borate buffer, pH 8.5. Change the buffer twice over a 10 min period.
4. Wash slides with PBS, changing the solution three times over a 10 min period.
5. Place slides in a humidified chamber (e.g., a sealed plastic box layered with wet paper towels) and cover cells with 150-300 μL of solution containing approximately 6 μg/mL anti-bromodeoxyuridine antibody diluted in PBS with 0.1% BSA. Incubate for 60 min at room temperature.
6. Wash slides with PBS, changing the solution three times over a 10 min period.
7. Apply optimal dilution of a second antibody conjugate (e.g., anti-mouse IgG-peroxidase), incubate, wash, and perform detection with a substrate that produces an insoluble product. After detection, counterstain with Harris-modified hematoxylin if desired. Slides can then be dehydrated and mounted.生物信息 免疫原品种 Bromodeoxyuridine-bovine serum albumin concentrate 克隆 AH4H7-1 / 131-14871 浓缩 Please refer to the Certificate of Analysis for the lot-specific concentration. 宿主 Mouse 特异性 Binds to bromodeoxyuridine and crossreacts with iodouridine (10%). Anti-bromodeoxyuridine does not crossreact with fluorodeoxyuridine, nor with any endogenous cellular components such as thymidine or uridine. 同种型 IgG1 品种反应性 All 抗体类型 Monoclonal Antibody 纯化方法 Protein A purfied 产品使用声明 使用声明 - 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.
储存和货运信息 存储条件 Maintain for 6 months 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. 包装信息 数量 50 µg
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