Key features and details
- One-wash 90 minute protocol
- Sensitivity: 50 pg/ml
- Range: 390.63 pg/ml - 25000 pg/ml
- Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Milk, Saliva, Serum
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Product nameHuman IgM ELISA Kit
See all IgM kits
Intra-assay Sample n Mean SD CV% Plasma 8 4.9% Inter-assay Sample n Mean SD CV% Plasma 3 5.8%
Sample typeCell culture supernatant, Saliva, Milk, Serum, Hep Plasma, EDTA Plasma, Cit plasma
Assay typeSandwich (quantitative)
Range390.63 pg/ml - 25000 pg/ml
Sample specific recovery Sample type Average % Range Cell culture supernatant 100 94% - 105% Saliva 101 94% - 110% Milk 96 90% - 101% Serum 96 93% - 98% Hep Plasma 98 97% - 100% EDTA Plasma 102 101% - 103% Cit plasma 108 104% - 114%
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Human
Does not react with: Goat, Cow
Human IgM ELISA Kit (ab214568) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of IgM protein in cell culture supernatant, cit plasma, edta plasma, hep plasma, milk, saliva, and serum. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human IgM with 50 pg/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 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) = 0.000249 (2.5e-4 IU) x SimpleStep IgA value (ng/mL)
Immunoglobulin M, or IgM, is a basic antibody produced by B cells and the largest of the immunoglobulins in the Human circulatory system. IgM typically constitutes about 10% of serum immunoglobulins and is prominent in the early immune responses. IgM (with IgD) is the major immunoglobulin expressed on the surface of B cells. IgM typically forms pentamers which includes the J chain, while in the less common hexameric form the J chain is absent. Although the common pentameric form of IgM contains 10 antigen binding sites, IgM cannot bind 10 antigens simultaneously due to size constraints. Class specific anti-immunoglobulin antibodies are useful for the characterization of malignant B cell proliferations. Most lymphocytic leukemias share either surface or intra cytoplasmic Ig with an isotypic restriction, which suggest the monoclonal nature of the cell population. Most of the chronic lymphocytic leukemias, non Hodgkin lymphomas and Burkitt's lymphoma bear surface IgM, whereas plasmocytes from Waldenström's disease bear intracytoplasmic IgM.
PlatformPre-coated microplate (12 x 8 well strips)
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent 5BC 1 x 6ml 10X Human IgM Capture Antibody 1 x 600µl 10X Human IgM Detector Antibody 1 x 600µl Human IgM Lyophilized Purified Protein 2 vials Plate Seals 1 unit SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Sample Diluent NS (ab193972) 1 x 50ml Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml
RelevanceIgM normally constitutes about 10% of serum immunoglobulins. IgM antibody is prominent in early immune responses to most antigens and predominates in certain antibody responses such as 'natural' blood group antibodies. IgM (with IgD) is the major immunoglobulin expressed on the surface of B cells. The gene for the mu constant region contains four domains separated by short intervening sequences. Class specific anti immunoglobulin antibodies are useful for: The characterization of malignant B cell proliferations. All but acute lymphocytic leukemias share either surface or intra cytoplasmic Ig with an isotypic restriction, which suggest the monoclonal nature of the cell population. Most of the chronic lymphocytic leukemias, non Hodgkin lymphomas and Burkitt's lymphoma bear surface IgM, whereas plasmocytes from Waldenström's disease bear intracytoplasmic IgM. The other isotypes are less frequently found. On the other hand multiple myelomas are usually of the IgG or IgA type. Characterization of plasma cells in inflammatory conditions: Plasma cell typing can be of use for the classification of intestinal inflammatory conditions such as inflammatory bowel disease and allergic conditions. In the latter a specific increase in the number of IgE plasma cells can be demonstrated.
Cellular localizationIsoform 1: Secreted. During differentiation, B lymphocytes switch from expression of membrane bound IgM to secretion of IgM. Isoform 2: Cell membrane; Single pass type I membrane protein.
- Constant region of heavy chain of IgM
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 IgM were measured in duplicates, interpolated from the IgM standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:40,000, plasma (citrate) 1:40,000, plasma (EDTA) 1:40,000 and plasma (heparin) 1:40,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IgM concentration was determined to be 618.9 µg/mL in neat serum, 559.0 µg/mL in neat plasma (citrate), 705.1 µg/mL in neat plasma (EDTA), and 618.2 µg/mL in neat plasma (heparin).
The concentrations of IgM were measured in duplicates, interpolated from the IgM standard curves and corrected for sample dilution. Undiluted samples are as follows: milk 1:750, saliva 1:400, and PBMC cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IgM concentration was determined to be 14,328 ng/mL in neat milk, 7,869 ng/mL in neat saliva and 16.0 ng/mL in neat PBMC cell culture supernatant.
Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IgM concentration was determined to be 524 µg/mL with a range of 178 – 901 µg/mL.
ab214568 has been referenced in 1 publication.
- Jindan RA et al. Clinical Interpretation of Detection of IgM Anti-Brucella Antibody in the Absence of IgG and Vice Versa; a Diagnostic Challenge for Clinicians. Pol J Microbiol 68:51-57 (2019). PubMed: 31050253