Overview

  • Product name

    Protein A ELISA Kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 3.7%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 10.2%
  • Sample type

    Cell culture supernatant
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    3.7 pg/ml
  • Range

    7.8 pg/ml - 500 pg/ml
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Product overview

    Protein A in vitro SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Protein A protein in purified IgG preparations and cell culture supernatant. 


    The SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Protein A (Staphylococcus aureus Protein A) is a 42 kDa protein that is characterized by its ability to bind the Fc portion of immunoglobulins.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Protein A Capture Antibody 1 x 600µl
    10X Protein A Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BC 1 x 6ml
    Plate Seals 1 unit
    Protein A Lyophilized Recombinant Protein 2 vials
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Relevance

    Protein A is a surface protein of S.aureus which binds IgG molecules by their Fc region. In serum, the bacteria will bind IgG molecules in the wrong orientation on their surface which hinders opsonization and phagocytosis. Mutants of S. aureus lacking protein A are more efficiently phagocytosed in vitro, and mutants in infection models have diminished virulence. Due to its affinity for the Fc region of many mammalian immunoglobulins, protein A is considered a universal reagent in biochemistry and immunology.
  • Cellular localization

    Secreted; cell wall.
  • Alternative names

    • gG-binding protein A
    • Immunoglobulin G binding protein A
    • SPA
    • Staphylococcal protein A

Applications

Our Abpromise guarantee covers the use of ab215542 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Protein A were measured in duplicate and interpolated from the Protein A standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Protein A concentration was determined to be 442 pg/mL in the Rabbit Monoclonal Antibody sample.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Protocols

References

ab215542 has not yet been referenced specifically in any publications.

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