Overview

  • Product nameIgG Human ELISA KitSee all IgG kits ...
  • Detection methodColorimetric
  • Tests
    1 x 96 well plate
  • Sample type
    Serum, Plasma
  • Assay typeSandwich (quantitative)
  • Sensitivity
    < 150 pg/ml
  • Range
    0.021 ng/ml - 15 ng/ml
  • Recovery

    117 %

    Sample specific recovery
    Sample type Average % Range
    Serum 116.1 105% - 123%
    Plasma 118.1 99% - 126%

  • Assay durationMultiple steps standard assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Abcam’s IgG Human ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human IgG in serum and plasma.

    This assay employs an antibody specific for Human IgG coated on a 96-well plate. Standards and samples are pipetted into the wells and IgG present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human IgG antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IgG bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Notes

    The Human immune system consists of two functional components classified as the innate system (the physical, biochemical and cellular barriers), and the adaptive immune system (including lymphocytes and immunoglobulins). Immunoglobulins are the key elements of the humoral immune response in vertebrate against parasitic invasion.

    The polypeptide chains of immunoglobulins composed of two identical heavy (H) chains and two identical light (L) chains linked together by inter-chain disulfide bonds. While the amino-terminal portions that exhibits highly variable amino-acid composition are involved in antigen binding, the C terminal constant parts are involved in complement binding, placental passage and binding to cell membranes. Based upon the variation of the constant region of the heavy chain, nine immunoglobulin heavy chain isotypes are found in humans: IgA (with subclasses IgA1 and IgA2), IgD, IgE, IgM, and IgG (with subclasses IgG1, IgG2, IgG3, and IgG4).

    IgG is the predominant immunoglobulin in the serum, which accounts for 75% of the total serum antibody of healthy individuals. IgG has a molecular weight of about 150 kDa. Four distinct subgroups of human IgG (IgG1, IgG2, IgG3, and IgG4) were first demonstrated in the 1960’s by using polyclonal antisera prepared in animals immunized with human myeloma proteins.

    The capture antibody of the kit ab100547 detects the Fc portion of the heavy chain of all subclasses of human IgG and the detection antibody detects the F(ab’)2 fragment of IgG.

  • Tested applicationsSandwich ELISAmore details
  • PlatformMicroplate

Properties

  • RelevanceImmunoglobulin G (IgG), is one of the most abundant proteins in human serum with normal levels between 8-17 mg/ml in adult blood. IgG is important for our defence against microorganisms and the molecules are produced by B lymphocytes as a part of our adaptive immune response. The IgG molecule has two separate functions; to bind to the pathogen that elicited the response and to recruit other cells and molecules to destroy the antigen. The variability of the IgG pool is generated by somatic recombination and the number of specificities in an individual at a given time point is estimated to be 1011 variants.
  • Cellular localizationCell Membrane and Secreted
  • Alternative names
    • Ig gamma
    • Ig gamma-1 chain C region
    • IGHG1
    • Immunoglobulin G
  • Database links

Applications

Our Abpromise guarantee covers the use of ab100547 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.

IgG Human ELISA Kit images

  • Hu IgG measured in biological fluids showing quantity (pg) per mL of tested sample
  • Representative standard curve using ab100547

Protocols

References for IgG Human ELISA Kit (ab100547)

ab100547 has not yet been referenced specifically in any publications.

Product Wall

I am sorry to read that the assay did not work. Were you able to get a standard curve, or were the absorbancevalues also low (compared to the values shown on the datasheet)?

In any case, I will replace the kit or refund the cost. The diluent...

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IgG levels in human milk smaples by ELISA

Average Average 3/5 (Ease of Use)
Abreviews
Abcam´s IgG human ELISA kit was performed according to manufacturer protocol except to sample preparation, because this protocol was not previously validated for human milk samples. Therefore, we had to dilute for achieve the expected concentrations in samples. For example, we did a dilution of 1: 1,000,000 to get 1-3 mg/mL concentrations (data from scientific studies) at follows:
Add 1 μL of serum human milk into a tube with 99 μL 1X Assay Diluent B to prepare a 100-fold diluted sample. Mix through and then pipette 1 μL of prepared 100-fold diluted sample into a tube with 99 μL 1X Assay Diluent to prepare a 10,000 fold diluted sample. Mix through and then pipette 1 μL of prepared 10,000-fold diluted sample into a tube with 99 μL 1X Assay
More optimized dilutions are required to improve the sensibility of the low concentrations. Moreover, it’s important to point out that these immunoglobulin levels are low in milk samples, unlike serum from blood. It would be advisable optimize the type of diluent and standards to this type of sample. Despite of these reasons, IgG was detected in all samples, showing levels between 0,200 and 500 mg/mL.
In our opinion, it would be necessary positive controls to perform the analysis.
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Verified customer

Submitted Aug 20 2014

1. I can confirm that the standard included with this kit is total human IgG purified from plasma

2. This kit can detect all IgG subclasses, however we have not determined the relative sensitivity to each.

Epitope mapping has not been performed. However, the capture antibody is a mouse monoclonal and recognizes the Fc portion and the detection antibody is polyclonal and recognizes the Fab portion.

Thanks for your enquiry.

Sample binds to an antibody specific for Human IgG coated on the 96-well plate.

I hope this is helpful. Please contact me again if you have any further questions.

The following kits are compatible as per your request;

http://www.abcam.com/iga-human-elisa-kit-ab137980.html

http://www.abcam.com/igg-human-elisa-kit-ab100547.html

http://www.abcam.com/igm-human-elisa-kit-ab137982.html

I would recommend use of a secondary antibody against mouse IgG which has been pre-absorbed against human IgG. These antibodies are run through columns containing IgGs of many species and and antibody which reacts with those IgGs will be trapped in the...

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It should in therory work. Unfortunately we do not have any information about the use of this kit in Mouse plasma. As there is no information available about the cross-reactivity of the antibody used in the kit with other species, I would recommen...

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Thank you for contacting us.

The standard of the kit is total human IgG purified from plasma. We do not sell kit components separately, however the equivalent in our catalogue would be ab98981, www.abcam.com/ab98981.

For the assay dil...

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Merci de nous avoir contactés.

Nous avons un kit ELISA qui mesure quantitativement le taux d’IgG humain dans le sérum et le plasma. La référence est ab100547et il coûte €490.00. Malheureusement, c...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"