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507858 PANSORBIN® Cells

PANSORBIN® Cells: Ficha de datos de seguridad (MSDS o SDS), certificado de análisis y de calidad (CoA y CoQ), expedientes, folletos y otros documentos disponibles.

Sinónimos: Staphylococcus aureus Cells

507858
Purchase on Sigma-Aldrich

Descripción

Replacement Information

Products

Número de referenciaEmbalaje Cant./Env.
507858-10GM Frasco de vidrio 10 gm
507858-1GM Frasco de vidrio 1 gm
507858-50GM Frasco de vidrio 50 gm
507858-5GM Ampolla de plást. 5 gm
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
Global Trade Item Number
Número de referencia GTIN
507858-10GM 04055977272505
507858-1GM 04055977272499
507858-50GM 04055977271942
507858-5GM 04055977271935

Documentation

PANSORBIN® Cells Ficha datos de seguridad (MSDS)

Título

Ficha técnica de seguridad del material (MSDS) 

PANSORBIN® Cells Certificados de análisis

CargoNúmero de lote
507858

Referencias bibliográficas

Visión general referencias
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.

Citas

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

    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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    Categorías

    Life Science Research > Protein Detection and Quantification > Immunodetection Support Products > Protein A & Protein G Reagents