A part of Merck

MAB3424 | Anti-BrdU Antibody, clone AH4H7-1 / 131-14871

MAB3424
50 µg  
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      Replacement Information

      Ofertas especiais

      Tabela com principais espec.

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      A FC, ICC, IHC M Purified Monoclonal Antibody
      Description
      Catalogue NumberMAB3424
      ReplacesMAB1467
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionAnti-BrdU Antibody, clone AH4H7-1 / 131-14871
      Alternate Names
      • BrdU
      Background InformationBromodeoxyuridine (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).
      References
      Product Information
      FormatPurified
      Control
      • After incorporation of BrdU, all DNA containing species
      PresentationLiquid in 10 mM Phosphate buffer, pH 7.4 containing 150 mM NaCl and 0.1% sodium azide
      Applications
      ApplicationUse Anti-BrdU Antibody, clone AH4H7-1 / 131-14871 (Mouse Monoclonal Antibody) validated in FC, ICC, IHC to detect Bromodeoxyuridine also known as BrdU.
      Key Applications
      • Flow Cytometry
      • Immunocytochemistry
      • Immunohistochemistry
      Application NotesImmunohistochemistry: (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.
      Biological Information
      ImmunogenBromodeoxyuridine-bovine serum albumin concentrate
      CloneAH4H7-1 / 131-14871
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostMouse
      SpecificityBinds 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.
      IsotypeIgG1
      Species Reactivity
      • All
      Antibody TypeMonoclonal Antibody
      Purification MethodProtein A Purfied
      Molecular Weightdependent upon the molecular weight of the bromodeoxyuridine incorporated protein being detected
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      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.
      Storage and Shipping Information
      Storage ConditionsMaintain 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.
      Packaging Information
      Material Size50 µg
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Título

      Ficha de Segurança de Produtos (MSDS) 

      Certificados de análise

      TítuloNúmero do lote
      Anti-Bromodeoxyuridine, clone AH4H7-1 / 131-14871 - 2420381 2420381
      Anti-Bromodeoxyuridine, clone AH4H7-1 / 131-14871 - 2145042 2145042
      Anti-Bromodeoxyuridine, -2519302 2519302
      Anti-Bromodeoxyuridine, -2637116 2637116
      Anti-Bromodeoxyuridine, -2661403 2661403
      Anti-Bromodeoxyuridine, -2736634 2736634
      Anti-Bromodeoxyuridine, -2802044 2802044
      Anti-Bromodeoxyuridine, clone AH4H7-1 / 131-14871 - 2302155 2302155
      Anti-Bromodeoxyuridine, clone AH4H7-1 / 131-14871 - NG1812620 NG1812620
      Anti-Bromodeoxyuridine, clone AH4H7-1 / 131-14871 - NG1861514 NG1861514
      Anti-Bromodeoxyuridine, clone AH4H7-1 / 131-14871 - NMM1701216 NMM1701216
      MOUSE ANTI-BROMODEOXYURIDINE (BrdU) MONOCLONAL ANTIBODY -

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      Life Science Research > Antibodies and Assays > Primary Antibodies