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507858 PANSORBIN® Cells

507858
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                  Description
                  Overview

                  PANSORBIN® Cells are heat-killed, formalin-fixed Staphylococcus aureus cells that have a coat of protein A and have been pickled by the method of Kessler. Useful for mitogenic stimulation of B lyphocytes and for immunoprecipitation.
                  Catalogue Number507858
                  Brand Family Calbiochem®
                  SynonymsStaphylococcus aureus Cells
                  References
                  ReferencesKierszenbaum, F., et al. 1991. Immunology 74, 317.
                  Meikle, P.J., et al. 1991. J. Biol. Chem. 266, 22569.
                  Ezaki, O., et al. 1989. Biochem. Biophys. Res. Commun. 159, 1368.
                  Murakami, H., et al. 1988. Biochem. J. 256, 917.
                  Kessler, S.W. 1975. J. Immunol. 115, 1617.
                  Product Information
                  Activity≥2.0 mg human IgG/100 mg cells
                  FormHomogeneous, milky suspension
                  FormulationSupplied as a 10% suspension of Staphylococcus aureus cells in PBS, 0.1% NaN₃, pH 7.2.
                  Quality LevelMQ100
                  Applications
                  Key Applications Immunoprecipitation
                  Biological Information
                  Physicochemical Information
                  ContaminantsViable cells: ≤2500 cells/ml
                  Dimensions
                  Materials Information
                  Toxicological Information
                  Safety Information according to GHS
                  Safety Information
                  Product Usage Statements
                  Storage and Shipping Information
                  Ship Code Ambient Temperature Only
                  Toxicity Standard Handling
                  Storage +2°C to +8°C
                  Do not freeze Yes
                  Packaging Information
                  Transport Information
                  Supplemental Information
                  Specifications

                  Documentation

                  PANSORBIN® Cells SDS

                  Title

                  Safety Data Sheet (SDS) 

                  PANSORBIN® Cells Certificates of Analysis

                  TitleLot Number
                  507858

                  References

                  Reference overview
                  Kierszenbaum, F., et al. 1991. Immunology 74, 317.
                  Meikle, P.J., et al. 1991. J. Biol. Chem. 266, 22569.
                  Ezaki, O., et al. 1989. Biochem. Biophys. Res. Commun. 159, 1368.
                  Murakami, H., et al. 1988. Biochem. J. 256, 917.
                  Kessler, S.W. 1975. J. Immunol. 115, 1617.

                  Citations

                  Title
                • Kira Steigerwald, et al. (2005) The APC tumor suppressor promotes transcription-independent apoptosis in vitro. Molecular Cancer Research 3, 78-89.
                • Data Sheet

                  Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

                  Revision06-May-2010 JSW
                  SynonymsStaphylococcus aureus Cells
                  DescriptionHeat-killed, formalin-fixed Staphylococcus aureus cells (Cowan I strain) that bear a high cell-surface density of protein A and have been pickled by the method of Kessler. Useful as a solid-phase IgG-binding reagent due to the high affinity interaction of protein A with the Fc domain of IgG. PANSORBIN® cells work best when the antibody is human (IgG1, IgG2, IgG4), rabbit IgG (all isotypes), or mouse (IgG2a, IgG2b, IgG3). Most common applications include immunoprecipitation and mitogenic stimulation of B lymphocytes.
                  FormHomogeneous, milky suspension
                  FormulationSupplied as a 10% suspension of Staphylococcus aureus cells in PBS, 0.1% NaN₃, pH 7.2.
                  Recommended reaction conditions
                  The procedures used for performing immunoprecipitations using PANSORBIN® cells are somewhat variable, but this protocol can serve as a general guideline. PANSORBIN® cells work best when the antibody is human (IgG1, IgG2, IgG4), rabbit IgG (all isotypes), or mouse (IgG2a, IgG2b, IgG3). Prewashing: 1. Transfer desired amount of PANSORBIN® cells to a microfuge tube (typically 10-30 µl of a 10% PANSORBIN® cell suspension per immunoprecipitation). 2. Centrifuge at 3000 x g for 5 min, or at maximum speed in a microfuge for 1 min at 4°C. 3. Aspirate the supernatant. 4. Resuspend to the original volume or in 1.0 ml of cold lysis buffer, whichever is greater. Mix gently. 5. Repeat wash (steps 2-4). 6. Resuspend the final pellet to the original volume (from step 1) in lysis buffer. 7. If kept sterile, washed PANSORBIN® cells are stable at 4°C. Pre-Clearing (Optional): Pre-clearing with PANSORBIN® cells prior to immunoprecipitation removes material that binds nonspecifically, reducing the background. 1. Add 10-30 µl of washed PANSORBIN® cells to the lysate prior to incubation with the primary antibody. 2. Incubate for 1 h on ice, mixing occasionally. 3. Centrifuge at 3000 x g for 5 min, or at maximum speed in a microfuge for 1 min at 4°C. 4. Transfer the supernatant to another microfuge tube and discard the pellet. Alternatively, the lysate can be pre-cleared with PANSORBIN® cells which have been pre-bound to nonspecific antiserum (see Secondary Antibody pre-binding procedure). Primary Antibody: 1. Add the required amount of primary antibody to the lysate. 2. Incubate for at least 1 h on ice, mixing occasionally. 3. If a secondary antibody step is not required or desired, proceed to the PANSORBIN® Step described below. Secondary Antibody: In some cases, a secondary antibody will be necessary. For example, if the primary antibody is mouse IgG1, it is best to use a rabbit anti-mouse IgG as a secondary antibody. If desired, the secondary antibody can be pre-bound to the PANSORBIN® cells to eliminate the loss of antibody-antigen complexes which have not bound to the protein A. 1. Incubate the desired amount of secondary antibody with prewashed PANSORBIN® cells for 45-60 min on ice. 2. Centrifuge at 3000 x g for 5 min, or at maximum speed in a microfuge for 1 min at 4°C. 3. Aspirate the supernatant and wash the lysis buffer. 4. Resuspend in lysis buffer to a volume equal to the original volume of PANSORBIN® cells which were added. Alternatively, the unbound secondary antibody and PANSORBIN® cells can be incubated with the sample sequentially, following primary antibody incubation. PANSORBIN® Step: 1. Add at least 10X the amount of PANSORBIN® cells (alone or bound to secondary antibody) needed to precipitate the primary (or secondary) antibody. PANSORBIN® cells bind about 2 mg of human IgG/ml. Typically, add 10-30 µl of PANSORBIN® cells per immunoprecipitation. This quantity gives a visible pellet. 2. Incubate for 1-2 h on ice, mixing regularly. 3. Centrifuge at 3000 x g for 5 min, or at maximum speed in a microfuge for 1 min at 4°C. 4. Wash PANSORBIN® pellet 2-3 times with lysis buffer. 5. If samples are to be loaded onto a polyacrylamide gel, the PANSORBIN® pellet can be resuspended in sample buffer, boiled for 10 min, centrifuged quickly (30 s in a microfuge at 20°C). Load the supernatant onto the gel. If a high background is observed, it may be necessary to pre-clear lysates, prebind the antibody to PANSORBIN® cells, or use a stronger lysis buffer (e.g. RIPA). This protocol is meant to serve as a guideline and may need to be modified for specific applications.
                  ContaminantsViable cells: ≤2500 cells/ml
                  Activity≥2.0 mg human IgG/100 mg cells
                  Storage +2°C to +8°C
                  Do Not Freeze Yes
                  Toxicity Standard Handling
                  ReferencesKierszenbaum, F., et al. 1991. Immunology 74, 317.
                  Meikle, P.J., et al. 1991. J. Biol. Chem. 266, 22569.
                  Ezaki, O., et al. 1989. Biochem. Biophys. Res. Commun. 159, 1368.
                  Murakami, H., et al. 1988. Biochem. J. 256, 917.
                  Kessler, S.W. 1975. J. Immunol. 115, 1617.
                  Citation
                • Kira Steigerwald, et al. (2005) The APC tumor suppressor promotes transcription-independent apoptosis in vitro. Molecular Cancer Research 3, 78-89.
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