Overview

  • 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, Heparin Plasma, EDTA Plasma, Citrate 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%
    Heparin Plasma 98 97% - 100%
    EDTA Plasma 102 101% - 103%
    Citrate Plasma 108 104% - 114%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat, Guinea pig, 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.

  • 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.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

  • 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 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Plate Seals 1 unit
  • Research areas
  • 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
    • DKFZp686I15212
    • FLJ00385
    • Hepatitis B virus receptor binding protein
    • Ig mu chain C region
    • IGHM
    • IgM heavy chain constant region
    • Immunoglobulin heavy constant mu
    • Immunoglobulin mu chain
    • Imunoglobulin heavy chain
    • Imunoglobulin heavy chain constant region mu
    • Imunoglobulin heavy chain mu constant region
    • MGC104996
    • MGC52291
    • MU
    • VH
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab229385 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • 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.

Protocols

References

ab229385 has not yet been referenced specifically in any publications.

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