BrdU Antibody | Bu20a

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A. BrdU-labeled human lymphoma cells were stained with Mouse anti BrdU antibody, clone BU20a, (MCA2483) at a 1/100 dilution overnight at 4°C. Goat anti Mouse IgG (H/L) FITC conjugated antibody (STAR117F) was used at a 1/50 dilution. READIDROP™ Propidium Iodide (1351101) was used to stain total DNA. B. BrdU-labeled human lymphoma cells were stained with Mouse anti BrdU antibody, clone BU20a, (MCA2483) at a 1/100 dilution for 1 hour at room temperature. Goat anti Mouse IgG (H/L) DyLight® 649 conjugated (STAR117D649GA) was used as the secondary detection reagent at a 1/50 dilution. PUREBLU™ Hoechst 33342 (1351304) was used to stain total DNA.

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  • Mouse anti BrdU
  • Mouse anti BrdU
  • Mouse anti BrdU
  • Mouse anti BrdU:FITC
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    BrdU antibody, clone Bu20a recognizes bromodeoxyuridine (known as BrdU or BrdUrd). This BU20a antibody is suitable for detecting incorporated BrdU in a wide variety of cell types and is suitable for use on tissue sections in double-labeling techniques.
    • Product Type
      Monoclonal Antibody
    • Clone
      Bu20a
    • Isotype
      IgG1
    2 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      MCA2483GAC, F *, Pdatasheet pdfdatasheet pdf0.1 mg
      MCA2483GA
      MCA2483C, F *, Pdatasheet pdfdatasheet pdf0.2 mg
      MCA2483
      MCA2483TC, F *, Pdatasheet pdfdatasheet pdf20 µg
      MCA2483T
      MCA2483FAF *datasheet pdfdatasheet pdf50 µg
      MCA2483FA
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
      -
      • Mouse anti BrdU antibody, clone Bu20a recognizes the thymidine analogue bromodoxyuridine (BrdU), which can be incorporated into DNA during S-phase of the cell cycle. The BU20a antibody is suitable for detecting incorporated BrdU in a wide variety of cell types and is suitable for use on tissue sections in double-labeling techniques (Makarev and Gorivodsky 2014).
      • Intended Use
      • Target Species
        Chemical
      • Product Form
        Purified IgG - liquid
        Purified IgG - liquid
        Purified IgG - liquid
        Purified IgG conjugated to Fluorescein Isothiocyanate Isomer 1 (FITC) - liquid
      • Reconstitution
      • Preparation
        Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
        Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
        Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
        Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
      • Preservative Stabilisers
        0.09%Sodium Azide (NaN3)
        0.09%Sodium Azide (NaN3)
        0.09%Sodium Azide (NaN3)
        0.09% Sodium Azide (NaN3)
        1% Bovine Serum Albumin
      • Immunogen
        Bromodeoxyuridine conjugated to BSA
      • Purity
      • Approx. Protein Concentrations
        IgG concentration 1.0 mg/ml
        IgG concentration 1.0 mg/ml
        IgG concentration 1.0 mg/ml
        IgG concentration 0.1 mg/ml
      • Reagents In The Kit
      • Preparing The Antibody
      • Test Principle
      • Buffer Solution
        Phosphate buffered saline
        Phosphate buffered saline
        Phosphate buffered saline
        Phosphate buffered saline
      • Fusion Partners
        Spleen cells from immunised Balb/c mice were fused with cells of the NS1 myeloma cell line
      • Storage
        Store at +4oC or at -20oC if preferred.
        Storage in frost-free freezers is not recommended.
        This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
        Store at +4oC or at -20oC if preferred.
        Storage in frost-free freezers is not recommended.
        This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
        Store at +4oC or at -20oC if preferred.
        Storage in frost-free freezers is not recommended.
        This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
        Store at +4oC or at -20oC if preferred.
        Storage in frost-free freezers is not recommended.
        This product should be stored undiluted. This product is photosensitive and should be protected from light.
        Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
      • Shelf Life
        18 months from date of despatch.
        18 months from date of despatch.
        18 months from date of despatch
        18 months from date of despatch.
      • Acknowledgements
      • Regulatory
        For research purposes only
      • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

      • Application NameYesNoMin DilutionMax Dilution
        Immunohistology - Frozen
        Immunohistology - Paraffin
        Flow Cytometry(1)1/251/100
        (1)
        See recommended protocol below.
      • Application NameYesNoMin DilutionMax Dilution
        Immunohistology - Frozen
        Immunohistology - Paraffin
        Flow Cytometry(1)1/251/100
        (1)
        See recommended protocol below.
      • Application NameYesNoMin DilutionMax Dilution
        Immunohistology - Frozen
        Immunohistology - Paraffin
        Flow Cytometry(1)1/251/100
        (1)
        See recommended protocol below.
      • Application NameYesNoMin DilutionMax Dilution
        Flow Cytometry(1)Neat1/10
        (1)
        See recommended protocol below.

      • Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.

      • Technical Advice
      • Technical Advice
      • Technical Advice
      • Technical Advice
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        1. BrdU label cells - add BrdU to the cell suspension in culture medium to a final concentration of 10 µM and incubate for 30 minutes in a CO2 incubator at 37°C

        2. Wash cells 2x with PBS/BSA, centrifuge cells at 500 x g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS

        3. Add cells slowly into 5 ml 70% ethanol chilled to -20°C, mix continuously by vortexing

        4. Incubate on ice for 30 minutes

        5. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend cell pellet

        6. Add 2 ml of 2 N HCl containing 0.5% Triton™ X-100, and incubate the cells for 30 minutes at room temperature on a rocking platform set to 15 rpm. Denaturation of the DNA by hydrochloric acid is critical for successful BrdU staining. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend in 1 ml of 0.1 M Na2B4O7, pH 8.5

        7. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend the cells in PBS/BSA + 0.05% Tween®-20 to a concentration of 1×107 cells/ml

        8. Aliquot 100 µl of the cell suspension into the required number of FACS tubes

        9. Incubate cells with mouse anti BrdU (Bu20a) antibody at the recommended dilution for 1hr at room temperature. Alternatively, incubation can be performed overnight at 4°C

        10. Wash step - add 2 ml of PBS/BSA, and centrifuge the cells at 500 x g for 5 minutes

        Optional Step: When using an unconjugated Bu20a antibody format perform incubation with a secondary antibody. For this purpose decant the supernatant and perform secondary antibody incubation for 45 minutes at room temperature (refer to the respective datasheet for information about the recommended dilution). Wash cells by repeating step 10 (wash step)

        11. Decant the supernatant and add 0.5 ml of sheath fluid

        12. For total DNA staining add READIDROP™ Propidium Iodide (1351101), READIDROP™ 7-AAD (1351102), or PUREBLU™ Hoechst 33342 (1351304) to each tube (refer to the respective datasheet for information about the recommended dilution)

        13. Analyze cells by flow cytometry following the manufacturer’s instructions. The DNA stain should be read on the appropriate channel set to the height and linear scale and not log scale
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        1. BrdU label cells - add BrdU to the cell suspension in culture medium to a final concentration of 10 µM and incubate for 30 minutes in a CO2 incubator at 37°C

        2. Wash cells 2x with PBS/BSA, centrifuge cells at 500 x g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS

        3. Add cells slowly into 5 ml 70% ethanol chilled to -20°C, mix continuously by vortexing

        4. Incubate on ice for 30 minutes

        5. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend cell pellet

        6. Add 2 ml of 2 N HCl containing 0.5% Triton™ X-100, and incubate the cells for 30 minutes at room temperature on a rocking platform set to 15 rpm. Denaturation of the DNA by hydrochloric acid is critical for successful BrdU staining. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend in 1 ml of 0.1 M Na2B4O7, pH 8.5

        7. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend the cells in PBS/BSA + 0.05% Tween®-20 to a concentration of 1×107 cells/ml

        8. Aliquot 100 µl of the cell suspension into the required number of FACS tubes

        9. Incubate cells with mouse anti BrdU (Bu20a) antibody at the recommended dilution for 1hr at room temperature. Alternatively, incubation can be performed overnight at 4°C

        10. Wash step - add 2 ml of PBS/BSA, and centrifuge the cells at 500 x g for 5 minutes

        Optional Step: When using an unconjugated Bu20a antibody format perform incubation with a secondary antibody. For this purpose decant the supernatant and perform secondary antibody incubation for 45 minutes at room temperature (refer to the respective datasheet for information about the recommended dilution). Wash cells by repeating step 10 (wash step)

        11. Decant the supernatant and add 0.5 ml of sheath fluid

        12. For total DNA staining add READIDROP™ Propidium Iodide (1351101), READIDROP™ 7-AAD (1351102), or PUREBLU™ Hoechst 33342 (1351304) to each tube (refer to the respective datasheet for information about the recommended dilution)

        13. Analyze cells by flow cytometry following the manufacturer’s instructions. The DNA stain should be read on the appropriate channel set to the height and linear scale and not log scale
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        1. BrdU label cells - add BrdU to the cell suspension in culture medium to a final concentration of 10 µM and incubate for 30 minutes in a CO2 incubator at 37°C

        2. Wash cells 2x with PBS/BSA, centrifuge cells at 500 x g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS

        3. Add cells slowly into 5 ml 70% ethanol chilled to -20°C, mix continuously by vortexing

        4. Incubate on ice for 30 minutes

        5. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend cell pellet

        6. Add 2 ml of 2 N HCl containing 0.5% Triton™ X-100, and incubate the cells for 30 minutes at room temperature on a rocking platform set to 15 rpm. Denaturation of the DNA by hydrochloric acid is critical for successful BrdU staining. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend in 1 ml of 0.1 M Na2B4O7, pH 8.5

        7. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend the cells in PBS/BSA + 0.05% Tween®-20 to a concentration of 1×107 cells/ml

        8. Aliquot 100 µl of the cell suspension into the required number of FACS tubes

        9. Incubate cells with mouse anti BrdU (Bu20a) antibody at the recommended dilution for 1hr at room temperature. Alternatively, incubation can be performed overnight at 4°C

        10. Wash step - add 2 ml of PBS/BSA, and centrifuge the cells at 500 x g for 5 minutes

        Optional Step: When using an unconjugated Bu20a antibody format perform incubation with a secondary antibody. For this purpose decant the supernatant and perform secondary antibody incubation for 45 minutes at room temperature (refer to the respective datasheet for information about the recommended dilution). Wash cells by repeating step 10 (wash step)

        11. Decant the supernatant and add 0.5 ml of sheath fluid

        12. For total DNA staining add READIDROP™ Propidium Iodide (1351101), READIDROP™ 7-AAD (1351102), or PUREBLU™ Hoechst 33342 (1351304) to each tube (refer to the respective datasheet for information about the recommended dilution)

        13. Analyze cells by flow cytometry following the manufacturer’s instructions. The DNA stain should be read on the appropriate channel set to the height and linear scale and not log scale
      • Recommended Protocol
        FLOW CYTOMETRY ANALYSIS

        1. BrdU label cells - add BrdU to the cell suspension in culture medium to a final concentration of 10 µM and incubate for 30 minutes in a CO2 incubator at 37°C

        2. Wash cells 2x with PBS/BSA, centrifuge cells at 500 x g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS

        3. Add cells slowly into 5 ml 70% ethanol chilled to -20°C, mix continuously by vortexing

        4. Incubate on ice for 30 minutes

        5. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend cell pellet

        6. Add 2 ml of 2 N HCl containing 0.5% Triton™ X-100, and incubate the cells for 30 minutes at room temperature on a rocking platform set to 15 rpm. Denaturation of the DNA by hydrochloric acid is critical for successful BrdU staining. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend in 1 ml of 0.1 M Na2B4O7, pH 8.5

        7. Centrifuge at 500 x g for 10 minutes, decant supernatant, and resuspend the cells in PBS/BSA + 0.05% Tween®-20 to a concentration of 1×107 cells/ml

        8. Aliquot 100 µl of the cell suspension into the required number of FACS tubes

        9. Incubate cells with mouse anti BrdU (Bu20a) antibody at the recommended dilution for 1hr at room temperature. Alternatively, incubation can be performed overnight at 4°C

        10. Wash step - add 2 ml of PBS/BSA, and centrifuge the cells at 500 x g for 5 minutes

        Optional Step: When using an unconjugated Bu20a antibody format perform incubation with a secondary antibody. For this purpose decant the supernatant and perform secondary antibody incubation for 45 minutes at room temperature (refer to the respective datasheet for information about the recommended dilution). Wash cells by repeating step 10 (wash step)

        11. Decant the supernatant and add 0.5 ml of sheath fluid

        12. For total DNA staining add READIDROP™ Propidium Iodide (1351101), READIDROP™ 7-AAD (1351102), or PUREBLU™ Hoechst 33342 (1351304) to each tube (refer to the respective datasheet for information about the recommended dilution)

        13. Analyze cells by flow cytometry following the manufacturer’s instructions. The DNA stain should be read on the appropriate channel set to the height and linear scale and not log scale
      • Flow Cytometry
        Use 10 μl of the suggested working dilution to label 1x106 cells in 100 μl
      • ELISA
      • ELISA
      • ELISA
      • ELISA
      • Immunohistology
      • Immunohistology
      • Immunohistology
      • Immunohistology
      • Histology Positive Control Tissue
      • Histology Positive Control Tissue
      • Histology Positive Control Tissue
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Immunofluorescence
      • Immunofluorescence
      • Immunofluorescence
      • Western Blotting
      • Western Blotting
      • Western Blotting
      • Western Blotting
      • Instructions For Use
      • Instructions For Use
      • Instructions For Use
      • Instructions For Use

      Additional BrdU Antibody Formats

      Formats Clone Applications Sizes available
      BrdU Antibody : Purified Bu20a C, F *, P 0.2 mg | 20 µg | 0.1 mg
      BrdU Antibody : FITC Bu20a F * 50 µg
      • Copyright © 2017 Bio-Rad Antibodies (formerly AbD Serotec)

      Recommended Secondary Antibody

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Goat anti Mouse IgG (H/L):DyLight®649 (Multi Species Adsorbed)STAR117D649GA0.1 mgF, IF
        STAR117D649GA
        Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed)STAR117F0.5 mgF
        STAR117F
        Rabbit F(ab')2 anti Mouse IgG:FITCSTAR9B1 mgF
        STAR9B
        Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed)STAR13B1 mgC, E, P, RE, WB
        STAR13B
        Rabbit F(ab')2 anti Mouse IgG:RPESTAR12A1 mlF
        STAR12A
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Goat anti Mouse IgG (H/L):DyLight®649 (Multi Species Adsorbed)STAR117D649GA0.1 mgF, IF
        STAR117D649GA
        Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed)STAR117F0.5 mgF
        STAR117F
        Rabbit F(ab')2 anti Mouse IgG:FITCSTAR9B1 mgF
        STAR9B
        Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed)STAR13B1 mgC, E, P, RE, WB
        STAR13B
        Rabbit F(ab')2 anti Mouse IgG:RPESTAR12A1 mlF
        STAR12A
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Goat anti Mouse IgG (H/L):DyLight®649 (Multi Species Adsorbed)STAR117D649GA0.1 mgF, IF
        STAR117D649GA
        Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed)STAR117F0.5 mgF
        STAR117F
        Rabbit F(ab')2 anti Mouse IgG:FITCSTAR9B1 mgF
        STAR9B
        Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed)STAR13B1 mgC, E, P, RE, WB
        STAR13B
        Rabbit F(ab')2 anti Mouse IgG:RPESTAR12A1 mlF
        STAR12A

        Recommended Negative Isotype Control

          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Mouse IgG1 Negative ControlMCA928100 TestsF
          MCA928
          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Mouse IgG1 Negative ControlMCA928100 TestsF
          MCA928
          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Mouse IgG1 Negative ControlMCA928100 TestsF
          MCA928

          Useful Reagents

            Recommended Positive Controls

              Histology Controls

                • Application NameReference Images
                  Immunofluorescence

                References

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                BrdU Staining Protocol

                Recommended guidance for staining with Mouse anti-BrdU antibody for flow cytometry and immunohistochemistry on paraffin-embedded tissue sections fixed in formalin.

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