Overview

  • Product name

    Human IgG ELISA Kit (10 x 96 well plate)
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 6.4%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 14.7%
  • Sample type

    Cell culture supernatant, Saliva, Milk, Urine, Serum, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    20 pg/ml
  • Range

    0.23 ng/ml - 15 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 89 80% - 106%
    Milk 89 83% - 94%
    Urine 87 82% - 93%
    Serum 101 88% - 125%
    Cell culture media 107 96% - 115%
    Hep Plasma 100 100% - 100%
    EDTA Plasma 90.17 87% - 93%
    Cit plasma 100 98% - 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, Cow, Dog
  • Product overview

    IgG in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IgG protein in human serum, plasma, milk, urine, saliva, culture media and tissue extracts.


    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. The levels of IgG are generally considered to be indicative of an individual’s immune status and are found increased in all types of infections, liver disease, severe malnutrition, dysproteinemia and rheumatoid arthritis. It is decrease in conditions such as hypogammaglobulinemia, X-linked agammaglobulinemia, lymphoid aplasia and chronic lymphoblastic leukemia. IgG accounts for 75% of the total human protein and can be found in serum, lymphatic fluid, cerebrospinal fluid, colostrum, milk, urine, saliva, sweat and body tissues. IgG has been shown to bind some bacterial strains from cutaneous microbiota.

    The Fc portion of human IgG is frequently used as the basis of prolonged pharmacokinetics as it is used as a fusion partner to extend the half-life of fusion proteins.

  • 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 10 x 96 tests
    10X Human IgG Capture Antibody 1 x 6ml
    10X Human IgG Detector Antibody 1 x 6ml
    10X Wash Buffer PT 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 5ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 50ml
    Antibody Diluent CP 1 x 60ml
    Human IgG Lyophilized Purified Protein 10 vials
    Plate Seals 10 units
    Sample Diluent NS 2 x 250ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 10 units
    Stop Solution 1 x 120ml
    TMB Development Solution 1 x 120ml
  • Research areas

  • Relevance

    IgG is a monomeric immunoglobulin, built of two heavy chains gamma and two light chains. Each molecule has two antigen binding sites. This is the most abundant immunoglobulin and is approximately equally distributed in blood and in tissue liquids, constituting 75% of serum immunoglobulins in humans. This is the only isotype that can pass through the human placenta, thereby providing protection to the fetus in its first weeks of life before its own immune system has developed. It can bind to many kinds of pathogens, for example viruses, bacteria, and fungi, and protects the body against them by complement activation (classic pathway), opsonization for phagocytosis and neutralisation of their toxins. There are 4 subclasses: IgG1 (66%), IgG2 (23%), IgG3 (7%) and IgG4 (4%).
  • Cellular localization

    Cell Membrane and Secreted
  • Alternative names

    • FLJ39988
    • FLJ40036
    • FLJ40253
    • FLJ40587
    • FLJ40789
    • FLJ40834
    • G1m marker
    • G2m marker
    • G3m marker
    • G4m marker
    • HDC
    • Heavy chain disease protein
    • Human immunglobulin G
    • Ig gamma 1 chain C region
    • Ig gamma 2 chain C region
    • Ig gamma 3 chain C region
    • Ig gamma 4 chain C region
    • IgG heavy chain locus
    • IGHG 1
    • IGHG 2
    • IGHG 3
    • IGHG 4
    • IGHG1
    • IGHG2
    • IGHG3
    • IGHG4
    • Immunoglobulin Gm1
    • Immunoglobulin Gm3
    • Immunoglobulin Gm4
    • Immunoglobulin heavy constant gamma 1
    • immunoglobulin heavy constant gamma 1 (G1m marker)
    • Immunoglobulin heavy constant gamma 2
    • immunoglobulin heavy constant gamma 2 (G2m marker)
    • Immunoglobulin heavy constant gamma 3
    • immunoglobulin heavy constant gamma 3 (G3m marker)
    • Immunoglobulin heavy constant gamma 4
    • immunoglobulin heavy constant gamma 4 (G4m marker)
    • MGC117419
    • MGC45809
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab212169 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.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Ten individual healthy donors were evaluated for the presence of IgG in serum using this assay. Results were interpolated from the standard curve in Sample Diluent NS and corrected for sample dilution (1:5x106). The mean level of IgG was found at 12.5 mg/mL with a range of 9 – 18.4mg/mL.

  • Bodily fluids from 3 different donors were evaluated for the presence of IgG using this assay. Results were interpolated from the standard curve in sample diluent NS and corrected for sample dilution (1:2.5x104). The mean levels in Milk were found at 20.7 µg/mL, in Urine at 0.8 µg/mL and in Saliva at 11.1 µg/mL.

Protocols

References

ab212169 has not yet been referenced specifically in any publications.

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