Leucoperm™

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Peripheral dog granulocytes stained with Mouse anti Dog CD107b:RPE (MCA2558PE) following permeabilisation with Leucoperm® (BUF09)

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  • Product Type
    Accessory Reagent
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    BUF09CFdatasheet pdfdatasheet pdf datasheet pdf1000 Tests
    BUF09C
    BUF09BFdatasheet pdfdatasheet pdf datasheet pdf250 Tests
    BUF09B
    BUF09Fdatasheet pdfdatasheet pdf datasheet pdf50 Tests
    BUF09
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Flow cytometric analyses with monoclonal antibodies have been restricted primarily to cell surface molecules. Intracellular structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, immunoglobulins etc. were largely excluded from such studies.

      Also excluded from flow cytometric studies were cytoplasmic localisations of well established membrane molecules such as CD3 and CD22.

      LEUCOPERM reagents allow intracellular antigen analysis with the same ease as surface antigens. The only prerequisite is the availability of suitable antibody conjugates. Most commercially available monoclonal antibody conjugates can be used with LEUCOPERM reagents. Some determinants are sensitive, however, to the fixation step involved. This and the optimal fixation time may have to be determined experimentally for each antibody conjugate.
    • Intended Use
    • Product Form
      Reagent A - Fixation medium
      Reagent B - Permeabilisation medium
      Reagent A - Fixation medium
      Reagent B - Permeabilisation medium
      Reagent A - Fixation medium
      Reagent B - Permeabilisation medium
    • Reconstitution
    • Preparation
    • Preservative Stabilisers
      Formaldehyde in Reagent A
      Formaldehyde in Reagent A
      Formaldehyde in Reagent A
    • Purity
    • Approx. Protein Concentrations
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
    • Storage
      LEUCOPERM Cell Permeabilisation reagents should be stored and used at room temperature. DO NOT FREEZE. Do not use reagents if a precipitate forms or discolouration occurs.
      LEUCOPERM Cell Permeabilisation reagents should be stored and used at room temperature. DO NOT FREEZE. Do not use reagents if a precipitate forms or discolouration occurs.
      LEUCOPERM Cell Permeabilisation reagents should be stored and used at room temperature. DO NOT FREEZE. Do not use reagents if a precipitate forms or discolouration occurs.
    • Shelf Life
      12 months from date of despatch.
      12 months from date of despatch.
      12 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
      Flow Cytometry
    • Application NameYesNoMin DilutionMax Dilution
      Flow Cytometry
    • Application NameYesNoMin DilutionMax Dilution
      Flow Cytometry


    • LEUCOPERM reagents are intended for fixing cells in suspension with Reagent A and then permeabilising the cells with Reagent B. This procedure gives antibodies access to intracellular structures and leaves the morphological scatter characteristics of the cells intact. Specific formulations reduce background staining and allow simultaneous addition of permeabilisation medium and fluorochrome labelled antibodies.
    • Technical Advice
    • Technical Advice
    • Technical Advice
    • Recommended Protocol
    • Recommended Protocol
    • Recommended Protocol
    • ELISA
    • ELISA
    • ELISA
    • Immunohistology
    • Immunohistology
    • Immunohistology
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Immunofluorescence
    • Immunofluorescence
    • Western Blotting
    • Western Blotting
    • Western Blotting
    • Instructions For Use
      For the detection of cell cycle antigens such as Ki-67, PCNA and BrdU, methanol modification is recommended - see protocol #F5.

      1. Prepare cells in the appropriate manner. Adjust cell suspension to a concentration of 1 x 107 cells/ml in PBS/BSA. Whole blood samples may also be used. Bio-Rad recommend the use of EDTA anti-coagulant in these circumstances, although satisfactory results may be obtained using heparin or acid-citrate dextrose.

      2. Add 100ul of cell suspension to the appropriate number of test tubes.
      If required, perform staining of cell surface antigens at this stage. Following staining for the recommended period, wash cells once in PBS/BSA and discard supernatant.

      3. Add 100ul of Reagent A (fixation medium, stored at room temperature).

      4. Incubate for 15 minutes at room temperature.

      5. Add 3ml PBS/BSA and centrifuge for 5 minutes at 300 x g. Remove supernatant.

      6. Resuspend cells in 100ul of Reagent B (Permeabilization Medium).

      7. Immediately add recommended volume of the appropriate directly conjugated antibody. Vortex and incubate for 30 minutes at room temperature.
      If using an unconjugated primary antibody, wash in 3ml of PBS/BSA (as per step 5) and then repeat step 7 using an appropriate secondary antibody. There is no requirement to add further Leucoperm.

      8. Wash once in PBS/BSA. Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.25ml of 0.5% formaldehyde and store them at 2-8oC in the dark. Analyse fixed cells within 24 hours.

    • Instructions For Use
      For the detection of cell cycle antigens such as Ki-67, PCNA and BrdU, methanol modification is recommended - see protocol #F5.

      1. Prepare cells in the appropriate manner. Adjust cell suspension to a concentration of 1 x 107 cells/ml in PBS/BSA. Whole blood samples may also be used. Bio-Rad recommend the use of EDTA anti-coagulant in these circumstances, although satisfactory results may be obtained using heparin or acid-citrate dextrose.

      2. Add 100ul of cell suspension to the appropriate number of test tubes.
      If required, perform staining of cell surface antigens at this stage. Following staining for the recommended period, wash cells once in PBS/BSA and discard supernatant.

      3. Add 100ul of Reagent A (fixation medium, stored at room temperature).

      4. Incubate for 15 minutes at room temperature.

      5. Add 3ml PBS/BSA and centrifuge for 5 minutes at 300 x g. Remove supernatant.

      6. Resuspend cells in 100ul of Reagent B (Permeabilization Medium).

      7. Immediately add recommended volume of the appropriate directly conjugated antibody. Vortex and incubate for 30 minutes at room temperature.
      If using an unconjugated primary antibody, wash in 3ml of PBS/BSA (as per step 5) and then repeat step 7 using an appropriate secondary antibody. There is no requirement to add further Leucoperm.

      8. Wash once in PBS/BSA. Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.25ml of 0.5% formaldehyde and store them at 2-8oC in the dark. Analyse fixed cells within 24 hours.

    • Instructions For Use
      For the detection of cell cycle antigens such as Ki-67, PCNA and BrdU, methanol modification is recommended - see protocol #F5.

      1. Prepare cells in the appropriate manner. Adjust cell suspension to a concentration of 1 x 107 cells/ml in PBS/BSA. Whole blood samples may also be used. Bio-Rad recommend the use of EDTA anti-coagulant in these circumstances, although satisfactory results may be obtained using heparin or acid-citrate dextrose.

      2. Add 100ul of cell suspension to the appropriate number of test tubes.
      If required, perform staining of cell surface antigens at this stage. Following staining for the recommended period, wash cells once in PBS/BSA and discard supernatant.

      3. Add 100ul of Reagent A (fixation medium, stored at room temperature).

      4. Incubate for 15 minutes at room temperature.

      5. Add 3ml PBS/BSA and centrifuge for 5 minutes at 300 x g. Remove supernatant.

      6. Resuspend cells in 100ul of Reagent B (Permeabilization Medium).

      7. Immediately add recommended volume of the appropriate directly conjugated antibody. Vortex and incubate for 30 minutes at room temperature.
      If using an unconjugated primary antibody, wash in 3ml of PBS/BSA (as per step 5) and then repeat step 7 using an appropriate secondary antibody. There is no requirement to add further Leucoperm.

      8. Wash once in PBS/BSA. Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.25ml of 0.5% formaldehyde and store them at 2-8oC in the dark. Analyse fixed cells within 24 hours.

    Additional Leucoperm™ Formats

    Formats Applications Sizes available
    Leucoperm™ : Reagent F 1000 Tests | 250 Tests | 50 Tests
    • Copyright © 2016 Bio-Rad

    Recommended Secondary Antibody

      Recommended Negative Isotype Control

        Useful Reagents

          Recommended Positive Controls

            Histology Controls

              References

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