Human IgM ELISA Kit, Fluorescent (ab229385)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 32 pg/ml
- Range: 48.8 pg/ml - 200000 pg/ml
- Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Milk, Saliva, Serum
- Detection method: Fluorescent
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human IgM ELISA Kit, Fluorescent
See all IgM kits -
Detection method
Fluorescent -
Precision
Intra-assay Sample n Mean SD CV% Plasma 8 4.9% Inter-assay Sample n Mean SD CV% Plasma 3 5.8% -
Sample type
Cell culture supernatant, Saliva, Milk, Serum, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
32 pg/ml -
Range
48.8 pg/ml - 200000 pg/ml -
Recovery
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 time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human
Does not react with: Cow -
Product overview
IgM in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IgM protein in human serum, plasma, milk, saliva, 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
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.
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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 10X Human IgM Capture Antibody 1 x 600µl 10X Human IgM Detector Antibody 1 x 600µl Human IgM Lyophilized Purified Protein 2 vials Antibody Diluent 5BC 1 x 6ml 10X Wash Buffer PT (ab206977) 1 x 20ml Stoplight Red Substrate Buffer 1 x 12ml 100X Stoplight Red Substrate 1 x 120µl 500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated Black 96-Well Microplate 1 unit Plate Seals 1 unit -
Research areas
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Relevance
IgM 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 localization
Isoform 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. -
Alternative names
- AGM1
- Constant region of heavy chain of IgM
- DKFZp686I15196
see all -
Database links
- Entrez Gene: 3507 Human
- Omim: 147020 Human
- SwissProt: P01871 Human
Images
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Other - Human IgM ELISA Kit, Fluorescent (ab229385)
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 IgM standard curve in Sample Diluent NS.Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native IgM in human serum and plasma samples.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).
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Interpolated concentrations of native IgM in human milk, saliva, and PBMC cell culture supernatant samples.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.
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Serum from ten individual healthy human male donors was measured in duplicate.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.
