货号：MAB3424

品牌：Merck Millipore

规格：50Ug

目录价：￥5207.00

市场价格：￥4425.95

会员价格：￥4165.60

金山科研平台，产品价格货期咨询微信：jinshanbio Description: Anti-Bromodeoxyuridine Antibody, clone AH4H7-1 / 131-14871 | MAB3424 View All» Replaces: MAB1467 View All» Promotional Text: Special Shipping Offer on Antibodies100% Performance Guaranteed View All» Trade Name: Chemicon (Millipore) View All» Specificity: 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. View All» Molecular Weight: dependent upon the molecular weight of the bromodeoxyuridine incorporated protein being detected View All» Immunogen: Bromodeoxyuridine-bovine serum albumin concentrate View All» Clone: AH4H7-1 / 131-14871 View All» Isotype: IgG1 View All» Background Information: 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). View All» Species Reactivity: All View All» Application Notes: 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. View All» Control: After incorporation of BrdU, all DNA containing species View All» Purification Method: Protein A purfied View All» Presentation: Liquid in 10 mM Phosphate buffer, pH 7.4 containing 150 mM NaCl and 0.1% sodium azide View All» Storage Conditions: 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. View All» Alternate Names: BrdU View All» Usage Statement: 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. View All» Key Applications:

• Flow Cytometry

• Immunocytochemistry

• Immunohistochemistry
  View All» Brand Family: Chemicon View All» Product Name: Anti-Bromodeoxyuridine Antibody, clone AH4H7-1 / 131-14871 | MAB3424 View All» Concentration: 1.1 mg/mL View All» Antibody Type: Monoclonal Antibody View All» Qty/Pk: 50 μg View All» Format: Purified View All» Host: Mouse View All»
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