Overview

  • Product name
    Human IgA ELISA Kit, Fluorescent
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Serum 8 4.12%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 5.92%
  • Sample type
    Cell culture supernatant, Saliva, Milk, Urine, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    0.12 ng/ml
  • Range
    0.2 ng/ml - 100 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 90.31 87.91% - 92.45%
    Milk 92.21 81% - 101.51%
    Urine 99.48 86.21% - 109.34%
    Serum 98.37 81.35% - 109.17%
    Cell culture media 100.65 85.17% - 117.29%
    Heparin Plasma 97.17 91.05% - 109.18%
    EDTA Plasma 81.23 75.01% - 87.76%
    Citrate Plasma 93.47 88.78% - 98.99%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat, Rabbit, Goat, Guinea pig, Hamster, Cow, Dog, Pig
  • Product overview

    IgA in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IgA protein in humanserum, plasma, milk, saliva, urine, and cell culture supernatants.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint 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. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • Notes

    Human Immunoglobulin A (IgA) belongs to the immunoglobulin family of proteins which include Human IgG, IgE, and IgM, and are known as antibodies. IgA has an essential role in mucosal immunity. IgA is the second most common serum Ig at about one-fifth of the concentration of IgG, however it is the most abundant immunoglobulin in secretions, such as saliva, mucus, sweat, and tears. In normal human serum IgA is found mainly as a monomer, whereas in secretions IgA is found as a dimer. IgA dimeric form contains two additional proteins, secretory piece and a J chain. The secretory piece is synthesized in the epithelial cells and is added to IgA as it passes into the secretions, protecting it from degradation. Secretory IgA fights off bacteria by neutralizing the bacteria to clear the infection.

    The most common disease associated with high levels of IgA in the kidney is IgA nephropathy, also known as Berger’s disease. There is no clear clarification as to why IgA deposits occur in the kidney, however abnormality in the immune system is a possibility. Other diseases that IgA is associated with are Rheumatoid Arthritis, Crohn’s Disease, Celiac disease and Henoch-Schonlein purpura (HSP).

  • 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
    100X Stoplight Red Substrate 1 x 120µl
    10X Human IgA Capture Antibody 1 x 600µl
    10X Human IgA Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Antibody Diluent CPI 1 x 6ml
    Human IgA Lyophilized Purified Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
  • Research areas
  • Relevance
    Human IgA (immunoglobulin A) is a glycosylated protein of 160 kDa and is produced as a monomer or as a J chain linked dimer. Monomeric IgA constitutes 5-15 % of the serum immunoglobulins whereas dimeric IgA is localized to mucosa surfaces such as saliva, gastrointestinal secretion, bronchial fluids and milk. Mucosal IgA plays a major role in host defence by neutralising infectious agents at mucosal surfaces. The production is usually local and antigen specific IgA producing B cells can be found in regions under the lamina propria where they mature into dimeric IgA producing plasma cells. IgA deficiency is the most common immunodeficiency that may affect both serum and mucosal produced IgA. OR: The secretory component is a component of immunoglobulin A (IgA) which consists of a portion of the polymeric immunoglobulin receptor. Polymeric IgA binds to the polymeric immunoglobulin receptor on the basolateral surface of epithelial cells and is taken up into the cell via transcytosis. The receptor-IgA complex passes through the cellular compartments before being secreted on the luminal surface of the epithelial cells, still attached to the receptor. Proteolysis of the receptor occurs and the dimeric IgA molecule, along with the secretory component, are free to diffuse throughout the lumen.
  • Cellular localization
    Secreted
  • Alternative names
    • Hepatocellular carcinoma-associated protein TB6
    • Ig alpha 1 chain C region
    • Ig alpha 2 chain C region
    • IGHA
    • IGHA1
    • IGHA2
    • Immunoglobulin heavy constant alpha 1
    • Immunoglobulin heavy constant alpha 2
    • Immunoglobulin heavy constant alpha 2 A2m marker
    • PIgR
    • Poly-Ig receptor
    • polymeric immunoglobulin receptor
    • polymeric immunoglobulin receptor Secretory component
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab229396 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

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

  • Ten individual healthy donors were evaluated for the presence of IgA in serum using this assay. Results were interpolated from the standard curve in Sample Diluent NS and corrected for sample dilution (1:2X105). The mean level of Human IgA was found at 3.978 mg/mL with a range of 1.016 – 9.476 mg/mL.

  • Fluids from a pooled selection of healthy donors were evaluated for the presence of human IgA in serum, milk, saliva, and urine using this assay. Results were interpolated from the standard curve in Sample Diluent NS and corrected for sample dilution (1:2X105). The mean levels of human IgA in serum were found at 2.45 mg/mL, in milk at 0.74 mg/mL, in saliva at 0.94 mg/mL and in urine at 0.05 mg/mL.

Protocols

References

ab229396 has not yet been referenced specifically in any publications.

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