Overview

  • Product name

    Human IgG3 ELISA Kit
    See all IgG3 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Human Serum 8 3.73%
    Inter-assay
    Sample n Mean SD CV%
    Human Serum 3 4.01%
  • Sample type

    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    20 pg/ml
  • Range

    93.75 pg/ml - 6000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 93 86% - 102%
    Cell culture media 95 92% - 99%
    Heparin Plasma 84 81% - 89%
    EDTA Plasma 91 87% - 98%
    Citrate Plasma 91 81% - 102%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Sheep, Rabbit, Goat, Guinea pig, Hamster, Cow, Dog, Pig
  • Product overview

    Abcam’s IgG3 Human in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IgG3 protein in serum, plasma and culture supernatant.
    The 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. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Immunoglubulin G (IgG) is a glycoprotein molecule which belongs to the immunoglobulin family of proteins known as antibodies. Immunoglobulins are the key component of humoral immunity. IgG has an approximate molecular weight of about 150kDa and it is composed of four peptide chains: two identical heavy chains (γ) of about 50kDa and two identical light chains (κ) of about 25kDa each. The heavy chains are linked to each other and to the light chain by disulfide bonds. At the N terminus, both the heavy and the light chain contain variable regions (VH and VL) which account for antibody diversity. At the C terminus, both chains contain constant regions (CH and CL) but only CH mediates effector functions. Structurally the IgG molecule may be divided into: (1) the Fragment antigen binding region (Fab) containing the VL, VH, CL and CH2 all of which shape the antigen binding site and (2) the Fragment crystallizable region (Fc) containing CH domains 2 – 4 which stabilize the antibody and bind to the Fc receptor on the surface of macrophages, neutrophils, natural killer cells as well as to complement proteins to mediate therefore physiological effects.

    IgG is synthesized and secreted by plasma B cells in response to an immunogen after recognition of specific epitopes on the antigen and it is generated following class switching and maturation of an antibody response, thus providing immune protection. There are four subclasses of IgG in Humans (IgG 1, 2, 3, 4) with variable affinity to Fc receptors and complement. It has been shown that during HIV-1 infection, antibodies with the same epitope specificity but of different subclass have different functions.

    IgG3 comprises around 5 - 10% of total IgG and plays a major role in the immune responses against protein or polypeptide antigens. The affinity of the Fc region of IgG3 binds with very high affinity and mediates a strong antibody-dependent cellular cytotoxicity and complement dependent cytotoxicity. IgG3 does not bind to Protein A, but it does bind to Protein G and L. Antigen-specific IgG3 immune response has been associated with long-term protection from the Plasmodium falciparum parasite and Chikungunya virus and it has been shown to be correlated with a decreased HIV infection risk after vaccination induced immediate humoral response.

    IgG3 deficiency is associated with Wiscott-Aldrich disease, systemic lupus erythematosus, juvenile diabetes mellitus, and peridontal infections. IgG3 deficiency has been associated with a history of recurrent infections, leading to chronic lung disease

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human IgG3 Capture Antibody 1 x 600µl
    10X Human IgG3 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI 1 x 6ml
    Human IgG3 Lyophilized Purified Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Relevance

    IgG3, like IgG1, activates complement and has an affinity for fc receptors; thus IgG1 and IgG3 are the more effective of the IgG subclasses. An IgG3 deficiency is associated with Wiscott-Aldrich disease, systemic lupus erythematosus, juvenile diabetes mellitus, and peridontal infections. IgG anti-viral antibodies are restricted to IgG subclasses 1 and 3, with IgG3 antibodies being the first to appear in response to infection.
  • Alternative names

    • DKFZp686H11213
    • FLJ39988
    • FLJ40036
    • FLJ40253
    • FLJ40587
    • FLJ40789
    • FLJ40834
    • HDC
    • Heavy chain disease protein
    • Ig gamma 3 chain C region
    • IGHG3
    • Immunoglobulin Gm3
    • Immunoglobulin heavy constant gamma 3 (G3m marker)
    • MGC45809
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab201284 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

  • 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.

  • Native human IgG3 protein was measured in serum diluted 1:150,000 in sample diluent NS prior to a 2-fold dilution series in Sample Diluent NS. Native Human IgG3 protein was measured in citrate plasma (1:125,000), EDTA plasma (1:150,000), and heparin plasma (1:200,000) in a 2-fold dilution series in Sample Diluent NS. The interpolated dilution factor corrected values are graphed (mean +/- SD).

  • Ten individual healthy male donors and one pooled sample (n = 50) were evaluated for the presence of IgG3 in serum using this assay. Results were interpolated from the standard curve in Sample Diluent NS and corrected by sample dilution (1:200,000). In the 10 individual donors, the mean level of IgG3 is 381.44 µg/mL with a range of 173.71 – 746.69 µg/mL and a standard deviation of 159.71 µg/mL.

Protocols

References

ab201284 has not yet been referenced specifically in any publications.

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