Human IgG subclass ELISA kit

Name: Human IgG subclass ELISA kit
SKU: DEIA9474
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

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COA

SDS

Datasheet

Sample

serum

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Human IgG Subclass ELISA contains components required to construct an enzyme-linked immunoassay for the specific and quantitative measurement of Human IgG1, IgG2, IgG3, and IgG4 subclasses.

Contents of Kit

1. mAb Anti-Human IgG1, mAb Anti-Human IgG2, mAb Anti-Human IgG3, mAb Anti-Human IgG4: Storage: 2-8°C until expiration date. 4 vials × 2.5 mL.
2. Human Serum Control: 2 vials. Contains 0.1% sodium azide. Storage: 2-8°C until expiration date. Reconstitute the lyophilized control with Diluent Buffer.
3. Human IgG Subclass Standard: 2 vials. Contains 0.1% sodium azide. Storage: 2-8°C until expiration date.
4. Peroxidase Anti-Human IgG: 0.5 mL (50× concentrate). Storage: 2-8°C until expiration date.
5. TMB Solution: 25 mL
6. Stop Solution: 25 mL
7. Diluent Buffer: 135 mL
8. Wash Buffer Concentrate (25×): 100 mL.
9. Antibody-Coated Wells: 12 × 8 Well Strips, 2 Plates

Storage

2°C to 8°C

General Description

The Human IgG subclass ELISA Kit contains components required to construct an enzyme-linked immunoassay for the specific and quantitative measurement of Human IgG1, IgG2, IgG3, and IgG4 subclasses. Sufficient quantities of reagents are provided to yield 2 plates of 96 wells if the recommended assay procedure, storage and handling of materials are followed as specified on this insert. This kit is a sandwich-type ELISA using a horseradish peroxidase detection system. A coated microtiter plate captures monoclonal reagents that are specific to the various human IgG subclasses. The monoclonal antibodies in turn capture the human IgG subclasses, for which they are specific, out of the serum sample. These monoclonal antibodies have been characterized in an IUIS/WHO study. The captured human IgG is then labeled by a horseradish-peroxidase anti-human IgG reagent. The detection signal is then generated in proportion to the amount of human subclass antibody.

Citations

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Immunoglobulin G (IgG) is an essential component of the immune system, responsible for providing protection against pathogens. Within the IgG class, there are four distinct subclasses: IgG1, IgG2, IgG3, and IgG4. These subclasses exhibit variations in their structural properties and functional capabilities, contributing to the diverse roles they play in immune responses. Each IgG subclass has a unique distribution and abundance in the human body. IgG1 is the most prevalent subclass, accounting for approximately 66% of total IgG in the serum. IgG2 follows, representing around 23%, while IgG3 and IgG4 constitute smaller proportions, approximately 7% and 4%, respectively.

IgG and IgG subclasses structure. (Napodano C, 2021) Figure 1. Structure of IgG and of different IgG subclasses.
(Source: Napodano, C. et al., 2021)

The subclasses of IgG differ primarily in their constant regions, particularly in the hinges and upper CH2 domains. These regions are crucial for interactions with IgG-Fc receptors (FcγR) and C1q, enabling various effector functions. The ability to bind FcγR and C1q differs among the subclasses, leading to variations in their capacity to trigger immune cell responses and activate complement pathways. Furthermore, IgG subclasses exhibit variations in their ability to cross the placenta, impacting the transfer of maternal antibodies to the fetus during pregnancy. IgG1 efficiently crosses the placenta, providing passive immunity to the developing fetus. In contrast, IgG2 crosses the placenta to a lesser extent, while IgG3 and IgG4 show limited placental transfer.

The differences in effector functions, placental transfer, and other characteristics of IgG subclasses have significant implications in various clinical contexts. Monitoring the levels and ratios of IgG subclasses can aid in diagnosing and monitoring certain diseases, particularly immunodeficiencies and autoimmune disorders. Additionally, alterations in IgG subclass distribution have been observed in allergic diseases and chronic inflammatory conditions, providing insights into disease progression and treatment approaches.

Alternative Names

Human IgG subclass ELISA
Human IGHG1 subclass ELISA
Human Immunoglobulin G subclass ELISA
Human IGHG1 subclass ELISA kit
Human Immunoglobulin G subclass ELISA kit

References

1. Napodano C, et al. Immunological role of IgG subclasses. Immunological Investigations. 2021, 50(4): 427-444.

Datasheet

Certificate of Analysis

Safety Data Sheet

Q & A

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Q: Can we use this kit for Cerebrospinal Fluid analyses?

A: Regarding IgG Subclass Human ELISA Kit and its compatibility with CSF;
This kit may indeed be compatible with CSF however this is not a sample type we have explicitly tested for this kit. Thus, we can not guarantee compatibility. Having said that, many sample types should be compatible with most ELISAs but that sample will need to be optimized.This can be done by performing serial dilutions of your sample to ensure that the analyte of interest will fall into the optimal range to be detected, as well as to avoid things like Matrix Effects. Also helpful would be to perform a spike and recovery assay to determine if your sample medium is interfering with analyte detection.

Customer Reviews

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IgG subclasses and allotypes: From structure to effector functions

Frontiers in Immunology

Authors: Vidarsson, G., Dekkers, G., Rispens, T.

Abstract

Of the five immunoglobulin isotypes, immunoglobulin G (IgG) is most abundant in human serum. The four subclasses, IgG1, IgG2, IgG3, and IgG4, which are highly conserved, differ in their constant region, particularly in their hinges and upper CH2 domains. These regions are involved in binding to both IgG-Fc receptors (FcγR) and C1q. As a result, the different subclasses have different effector functions, both in terms of triggering FcγR-expressing cells, resulting in phagocytosis or antibody-dependent cell-mediated cytotoxicity, and activating complement. The Fc-regions also contain a binding epitope for the neonatal Fc receptor (FcRn), responsible for the extended half-life, placental transport, and bidirectional transport of IgG to mucosal surfaces. However, FcRn is also expressed in myeloid cells, where it participates in both phagocytosis and antigen presentation together with classical FcγR and complement. How these properties, IgG-polymorphisms and post-translational modification of the antibodies in the form of glycosylation, affect IgG-function will be the focus of the current review.

Immunoglobulin G (IgG) subclasses and human disease

The American Journal of Medicine

Authors: Oxelius, V. A.

Abstract

The isotypes of IgG, IgG1, IgG2, IgG3, and IgG4 were determined in immunoglobulin preparations and the effect on serum levels of treated patients. Serum IgG subclass deficiencies were recorded in different patient groups: (1) IgG2–IgG4 deficiency was associated with IgA deficiency. (2) IgG2–IgG4 deficiency was found in patients with ataxia telangiectasia. (3) Low IgG2 levels were recorded in patients with SLE; one of these patients with recurrent pericarditis was treated with immunoglobulin with good results. (4) Low IgG2 and/or low IgG3 levels were found in patients with juvenile diabetes mellitus. (5) Mothers giving birth to severely group B streptococci infected infants showed low levels of IgG subclasses indicating that the newborns were IgG subclass deficient at birth. (6) In a prospective study of children with recurrent otitis media aged 12 and 32 months the IgG2 levels were significantly reduced in the group with considerably high otitis proneness. In patients with IgG2–IgG4 deficiency, absence of antibodies to polysaccharide antigen teichoic acid and the protein antigen α-toxin of staphylococci was demonstrated. Imbalanced IgG subclass pattern with increased IgG4 was recorded in patients with different diseases such as atopic diseases and also in combination with increased IgE, Henoch-Sch?nlein vasculitis, idiopathic pulmonary hemosiderosis, chronic mucocutaneous candidiasis, and in patients with Trichuris trichurea infection.