Human IgM ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human IgM with a sensitivity of 32 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
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Key facts
- Detection method
- Fluorescent
- Sample types
- Saliva, Heparin Plasma, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 48.8 - 200000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 32 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
Constant region of immunoglobulin heavy chains. Immunoglobulins, also known as antibodies, are membrane-bound or secreted glycoproteins produced by B lymphocytes. In the recognition phase of humoral immunity, the membrane-bound immunoglobulins serve as receptors which, upon binding of a specific antigen, trigger the clonal expansion and differentiation of B lymphocytes into immunoglobulins-secreting plasma cells. Secreted immunoglobulins mediate the effector phase of humoral immunity, which results in the elimination of bound antigens (PubMed:20176268, PubMed:22158414). The antigen binding site is formed by the variable domain of one heavy chain, together with that of its associated light chain. Thus, each immunoglobulin has two antigen binding sites with remarkable affinity for a particular antigen. The variable domains are assembled by a process called V-(D)-J rearrangement and can then be subjected to somatic hypermutations which, after exposure to antigen and selection, allow affinity maturation for a particular antigen (PubMed:17576170, PubMed:20176268). Isoform 1. Constant region of secreted IgM (sIgM), also known as the Fc region of IgM antibody. Able to multimerize, forms high order polymers, mainly pentamers and occasionally hexamers, providing for multivalency and high avidity recognition of antigens (PubMed:32029689, PubMed:37095205). Natural sIgM are polyreactive and recognize conserved self- and pathogen-derived structures, whereas immune sIgM are secreted only upon exposure to pathogens and are antigen-specific. Both natural and immune sIgM are required for an efficient humoral immune response to infection (By similarity). Mediates sIgM effector functions mostly via Fc receptors and the complement system. On lymphoid cells binds high-affinity Fc receptor FCMR and promotes induction of an efficient neutralizing IgG response while maintaining tolerance to self-antigens. Recruits C1q complement component to initiate the classical complement pathway, facilitating the recognition and neutralization of pathogens by the host. Together with C1q and mannose-binding lectin promotes the phagocytosis of apoptotic cells by macrophages, ensuring the clearance of potential autoimmune epitopes from tissues (By similarity) (PubMed:12847249, PubMed:19006321, PubMed:28230186, PubMed:32029689). Involved in mucosal immunity. It is transported by transcytosis across mucosal epithelium by PIGR and secreted on the apical side in complex with PIGR secretory component to scan mucosal lining for pathogens. IgM-antigen complexes undergo FCMR-mediated retrotranscytosis across mucosal M cells toward antigen-presenting cells in mucosal lymphoid tissues (By similarity) (PubMed:32029689). Isoform 2. Constant region of membrane-bound IgM, part of the B cell receptor complex (BCR). IgM BCR provides constitutive tonic signaling for B cell survival. Mediates pre-BCR signaling that regulates B cell selection and rearrangement of Ig genes via allelic exclusion.
Alternative names
Immunoglobulin heavy constant mu, Ig mu chain C region, Ig mu chain C region BOT, Ig mu chain C region GAL, Ig mu chain C region OU, IGHM
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Recommended products
Human IgM ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human IgM with a sensitivity of 32 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Key facts
- Detection method
- Fluorescent
- Sample types
- Saliva, Heparin Plasma, Citrate plasma, Cell culture supernatant, Milk, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 48.8 - 200000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 32 pg/mL
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Plasma | n 8 | mean - | SD - | C.V. 4.9 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Plasma | n 3 | mean - | SD - | C.V. 5.8 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell culture supernatant | Average % = 100 | Range 94 - 105 % |
Sample type Milk | Average % = 96 | Range 90 - 101 % |
Sample type Serum | Average % = 96 | Range 93 - 98 % |
Sample type EDTA Plasma | Average % = 102 | Range 101 - 103 % |
Sample type Saliva | Average % = 101 | Range 94 - 110 % |
Sample type Heparin Plasma | Average % = 98 | Range 97 - 100 % |
Sample type Citrate plasma | Average % = 108 | Range 104 - 114 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
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.
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.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
IgM also known as immunoglobulin M is a type of antibody with a molecular mass of approximately 970 kDa. It is the largest antibody present in the human circulatory system mostly found in plasma and lymphatic tissue. IgM is secreted primarily by B cells as a pentamer with a J chain providing a high avidity for antigens. When bound to an antigen IgM can activate the complement system a part of the immune system that helps clear pathogens from an organism.
- Biological function summary
IgM acts as the first line of defense during the initial stages of an immune response. It plays an important part in agglutination and neutralization of pathogens forming a complex that alerts other components of the immune system to take action. IgM exists before class-switching to IgG providing immediate protection. It also plays a role in antibody-dependent cellular cytotoxicity. Although not typically a long-term solution its presence is critical for providing initial protection until more specific antibodies like IgG and IgA are produced.
- Pathways
The immune response that involves IgM begins with the activation of the classical complement pathway. This pathway bridges the innate and adaptive immunity ensuring the communication and coordination of immune responses. The pathway involves C1q protein binding to IgM-antigen complexes. IgM also interacts with antigen-presenting cells facilitating the processing of antigens and interaction with T cells. These interactions connect IgM with other proteins like IgG and IgA influencing various immune response pathways.
- Associated diseases and disorders
IgM is associated with conditions such as Waldenström's macroglobulinemia and rheumatoid arthritis. Waldenström's macroglobulinemia involves the overproduction of IgM by clonal B cells leading to symptoms like anemia bleeding and neuropathy. Rheumatoid arthritis shows increased IgM levels as part of the autoimmune response affecting joints. In these diseases the IgM antibody interacts with rheumatoid factor and other immune proteins involved in inflammation demonstrating its significant role in autoimmunity and abnormal immune responses.
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4 product images
Sandwich ELISA - Human IgM ELISA Kit, Fluorescent (ab229385)
Example of human IgM standard curve in Sample Diluent NS.
Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human IgM ELISA Kit, Fluorescent (ab229385)
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).
Sandwich ELISA - Human IgM ELISA Kit, Fluorescent (ab229385)
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.
Sandwich ELISA - Human IgM ELISA Kit, Fluorescent (ab229385)
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.
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