Human IgA ELISA Kit, Fluorescent (ab229396)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 0.12 ng/ml
- Range: 0.2 ng/ml - 100 ng/ml
- Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Milk, Saliva, Serum, Urine
- Detection method: Fluorescent
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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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, Hep Plasma, EDTA Plasma, Cit 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% Hep Plasma 97.17 91.05% - 109.18% EDTA Plasma 81.23 75.01% - 87.76% Cit 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: Goat, Cow, 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.
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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).
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Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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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 - HAMA Blocker (ab193969) 1 x 6ml Human IgA Lyophilized Purified Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated Black 96-Well Microplate 1 unit Stoplight Red Substrate Buffer 1 x 12ml -
Research areas
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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
see all -
Database links
- Entrez Gene: 3494 Human
- Entrez Gene: 5284 Human
- Omim: 146900 Human
- Omim: 147000 Human
- Omim: 173880 Human
- SwissProt: P01876 Human
- SwissProt: P01877 Human
Images
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Other - Human IgA ELISA Kit, Fluorescent (ab229396)
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.
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Example of human IgA standard curve in Sample Diluent NS.Background-subtracted data values (mean +/- SD) are graphed.
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IgA levels in individual healthy donors.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.
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Comparison of IgA levels in human serum, milk, saliva, and urine.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.
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Sandwich ELISA - Human IgA ELISA Kit, Fluorescent (ab229396)To learn more about the advantages of recombinant antibodies see here.
