Key features and details
- One-wash 90 minute protocol
- Sensitivity: 0.25 ng/ml
- Range: 0.78 ng/ml - 50 ng/ml
- Sample type: Cell culture supernatant, Milk, Plasma, Saliva, Serum, Urine
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Product nameHuman IgA ELISA Kit
See all IgA kits
Intra-assay Sample n Mean SD CV% Serum 8 4.12% Inter-assay Sample n Mean SD CV% Serum 3 5.92%
Sample typeCell culture supernatant, Saliva, Milk, Urine, Serum, Plasma
Assay typeSandwich (quantitative)
Range0.78 ng/ml - 50 ng/ml
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 time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Human
Does not react with: Goat, Cow, Pig
Human IgA ELISA Kit (ab196263) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of IgA protein in cell culture supernatant, milk, saliva, serum, urine, and plasma. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human IgA with 0.25 ng/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
This kit recognizes both native and purified Human IgA protein in serum, plasma, milk, urine, saliva, and cell culture media samples only.
Cell and tissue extracts have not been tested with this kit.
Human IgG, human IgM, and human IgE were prepared at 20 ng/mL and 10 ng/mL in Sample Diluent NS and assayed for cross reactivity. No significant cross reactivity was observed for human IgG, human IgM, or human IgE at either concentration with a mean OD deviation from background of 0.006.
Purified human IgA was assayed at 20 ng/mL in the presence and absence of 20 ng/mL of human IgM or human IgE to determine interference. After background subtraction, human IgM and IgE were found to interfere with the assay by 9.24%. Human IgA was assayed at 20 ng/mL in the presence and absence of 80 ng/mL of human IgG to determine interference. After background subtraction, no significant interference was observed.
This kit recognizes human IgA protein.
Other species reactivity was determined by measuring 1:200,000 (dilution) serum samples of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration in human serum assayed at the same dilution.
Reactivity < 3% was determined for the following species: Mouse, Rat, Hamster, Guinea Pig, Rabbit, Dog, Goat, Pig, Cow
This immunoassy is calibrated against a highly purified human IgA. The NIBSC/WHO unclassified purified human IgG/IgM/IgG preperation 67/086. was evaluatied in this kit.
The dose response curve of the unclassified standard parallels the SimpleStep standard curve. To Convert sample values obtained with the SimpleStep IgA kit to approximate NIBSC International units, use the equation below.
NIBSC 67/086 approximate value (IU/mL) = 0.000064 (6.4e-5 IU) x SimpleStep IgA value (ng/mL)
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 and is 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 monomeric, alternatively, in secretions, IgA is found as a dimer containing 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. IgA works is with lysozymes to hydrolyze carbohydrates in bacterial cells walls to clear the infection.
The most common disease associated with high levels of IgA in the kidney is IgA nephropathy, aka 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 Celiac disease and Henoch-Schonlein purpura (HSP).
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Human IgA Capture Antibody 1 x 600µl 10X Human IgA Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 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 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml
RelevanceHuman 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.
- Hepatocellular carcinoma-associated protein TB6
- Ig alpha 1 chain C region
- Ig alpha 2 chain C region
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.
Example of Human IgA standard curve in Sample Diluent NS. The Human IgA standard curve was prepared as described. Raw data values are shown in the table. 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.
Native IgA was measured in serum, plasma citrate, plasma EDTA, plasma heparin, milk, saliva and urine in a 2-fold dilution series. Sample dilutions were made in Sample Diluent NS. Purified IgA was spiked in culture media and diluted in a 2-fold dilution series in Sample Diluent NS.
Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.
Purified IgA was spiked in culture media and diluted in a 2-fold dilution series in Sample Diluent NS.
To learn more about the advantages of recombinant antibodies see here.
ab196263 has been referenced in 4 publications.
- Makridakis M et al. Multiplexed MRM-based protein quantification of putative prognostic biomarkers for chronic kidney disease progression in plasma. Sci Rep 10:4815 (2020). PubMed: 32179759
- Zhao J et al. Immunology, Treatment and Public Health Aspects of Subarachnoid Hemorrhage. Iran J Public Health 49:673-683 (2020). PubMed: 32548047
- Yoon JY et al. Aberrant expression of interleukin-10 and activation-induced cytidine deaminase in B cells from patients with Behçet's disease. Biomed Rep 7:520-526 (2017). PubMed: 29188055
- Zhou ZQ et al. Follicular helper T cell exhaustion induced by PD-L1 expression in hepatocellular carcinoma results in impaired cytokine expression and B cell help, and is associated with advanced tumor stages. Am J Transl Res 8:2926-36 (2016). PubMed: 27508013