Human IgG ELISA Kit (ab195215)
- Datasheet
- References (3)
- Protocols
Overview
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Product name
Human IgG ELISA Kit
See all IgG kits -
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, Hep Plasma, EDTA Plasma, Cit plasma, Tissue Lysate -
Assay type
Sandwich (quantitative) -
Sensitivity
0.02 ng/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, Rhesus monkey
Does not react with: Sheep, Goat, Cow -
Product overview
Human IgG ELISA kit (ab195215) is designed for the quantitative measurement of IgG protein in human serum, plasma, milk, urine, saliva, culture media and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human IgG with 20 pg/mL sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.
CROSS REACTIVITY Human IgM, human IgA and human IgE were prepared at 10 ng/mL and 250 ng/mL in Sample Diluent NS and assayed for cross reactivity. No cross-reactivity was observed for IgM or IgE at either concentration with a mean OD deviation from background of -0.01. No cross reactivity was observed for IgA at 10 ng/mL and only 0.4% cross reactivity at 250 ng/mL of IgA.
INTERFERENCE Purified human IgG was assayed at 5 ng/mL in the presence and absence of 250 ng/mL of human IgM, human IgA and human IgE to determine interference. After background subtraction, recovery of human IgG was observed at a mean of 98% with a standard deviation of 0.06.
SPECIES REACTIVITY This kit recognizes human IgG protein.
Other species reactivity was determined by measuring 10 ng/mL of purified IgG from various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration of human IgG assayed at the same concentration.
Reactivity < 3% was determined for the following species: Mouse, Rat, Rabbit, Dog, Goat, Sheep, Cow, Guinea Pig -
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.
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Platform
Microplate
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human IgG Capture Antibody 1 x 600µl 10X Human IgG 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 CP 1 x 6ml Human IgG Lyophilized Purified Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 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
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Cellular localization
Secreted -
Alternative names
- Ig gamma 1 chain C region
- Ig gamma 2 chain C region
- Ig gamma 3 chain C region
see all -
Database links
- Entrez Gene: 3500 Human
- Entrez Gene: 3501 Human
- Entrez Gene: 3502 Human
- Entrez Gene: 3503 Human
- SwissProt: P01857 Human
- SwissProt: P01859 Human
- SwissProt: P01860 Human
- SwissProt: P01861 Human
see all
Images
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Other - Human IgG ELISA Kit (ab195215)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.
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Example of Human IgG standard curve in Sample Diluent NS.Background-subtracted data values (mean +/- SD) are graphed.
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Raw data values for example of Human IgG standard curve in Sample Diluent NSRaw data values are shown in the table
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Example of Human IgG standard curve in 1X Cell Extraction Buffer PTR.Background-subtracted data values (mean +/- SD) are graphed.
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Raw data values for example of Human IgG standard curve in 1X Cell Extraction Buffer PTRRaw data values are shown in the table
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IgG levels in individual healthy donors.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.
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Comparison of IgG levels in Human milk, urine and saliva.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.
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IgG1-4 isotypes are detected by this kitHuman IgG1, IgG2, IgG3 and IgG4 were tested at 5ng/mL
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Linearity of dilution – native IgG in human serum, plasma (citrate, EDTA and Heparin), and culture mediaNative IgG was measured in the human serum, plasma citrate, plasma EDTA, plasma heparin and culture media in a 2-fold dilution series. Sample dilutions were made in Sample Diluent NS for all samples, purified IgG was spiked into culture media and diluted in a 2-fold dilution series in Sample Diluent NS
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Linearity of dilution – native IgG in human milk, urine, saliva and HLH extractNative IgG was measured in milk, saliva, urine and liver homogenate (HLH) in a 2-fold dilution series. Sample dilutions were made in Sample Diluent NS for all samples except for HLH, which was carried out in 1X cell extraction buffer PTR.
Protocols
References
This product has been referenced in:
- Adler AS et al. A natively paired antibody library yields drug leads with higher sensitivity and specificity than a randomly paired antibody library. MAbs 10:431-443 (2018). ELISA . Read more (PubMed: 29376776) »
- Akyol GY et al. IVIG activates Fc?RIIB-SHIP1-PIP3 Pathway to stabilize mast cells and suppress inflammation after ICH in mice. Sci Rep 7:15583 (2017). Read more (PubMed: 29138419) »
