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AB325733

Human IgG4 ELISA Kit, Chemiluminescent

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Human IgG4 ELISA Kit, Chemiluminescent is a 55-min single-wash SimpleStep Ignite™ ELISA for the rapid quantification of Human IgG4 with higher sensitivity and broader measuring range.

- Single 30 min incubation and single-wash step for results in less than an hour.
- Chemiluminescent detection allows a broad dynamic range: 13.72 - 30000 pg/ml with a sensitivity of 2.83 pg/mL.
- Design your own immunoassay: we also offer the conjugation-ready antibody pair.
- Patent pending

View Alternative Names

Immunoglobulin heavy constant gamma 4, Ig gamma-4 chain C region, IGHG4

7 Images
Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)
  • sELISA

Lab

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)

Standard curve comparison between the Chemiluminescent Human IgG4 SimpleStep ELISA and colorimetric Human IgG4 SimpleStep ELISA (ab320788) in 1X Wash Buffer PT. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)
  • sELISA

Lab

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)

Interpolated concentrations of human IgG4 in serum, plasma (citrate), plasma (EDTA), and plasma (heparin). Interpolated concentration of native IgG4 was measured in duplicate at different sample concentrations. Undiluted samples are 1 : 100,000 serum, plasma (citrate), plasma (EDTA), and plasma (heparin). The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Wash Buffer PT.

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)
  • sELISA

Lab

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)

Interpolated concentrations of IgG4 in normal human serum donors. Serum of ten individual healthy human female donors was measured in duplicate. Interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). The mean IgG4 concentration was determined to be 248 ug/mL with a range of 102 – 566 ug/mL.

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)
  • sELISA

Lab

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)

Dynamic range comparison between the Chemiluminescent Human IgG4 SimpleStep ELISA, colorimetric Human IgG4 SimpleStep ELISA (ab320788), and leading chemiluminescent competitor. The Chemiluminescent SimpleStep ELISA kit shows increased sensitivity and increased dynamic range.

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)
  • sELISA

Lab

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)

Cross reactivity of related immunoglobulin proteins. 50 ng/mL of natively purified human IgG1, IgG2, IgG3, IgA, IgD, IgE, and IgM were tested individually for cross-reactivity. Percent reactivity was determined by interpolating the protein concentration from the human IgG4 standard curve and expressing the interpolated concentration as a percentage of the protein concentration tested.

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)
  • sELISA

Lab

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)

Interpolated concentrations of human IgG4 in urine, saliva, and milk. Interpolated concentration of native IgG4 was measured in duplicate at different sample concentrations. Undiluted samples are as follows : urine 1 : 20, saliva 1 : 100, and milk 1 : 100. The interpolated dilution factor corrected values (to neat sample) are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Wash Buffer PT.

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)
  • sELISA

Lab

Sandwich ELISA - Human IgG4 ELISA Kit, Chemiluminescent (AB325733)

SimpleStep ELISA® Ignite technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 55 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.

Key facts

Detection method

Luminescent

Sample types

Serum, Citrate plasma, EDTA Plasma, Heparin Plasma, Cell culture media, Urine, Saliva, Milk

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 2.83 pg/mL

Range

13.72 - 30000 pg/mL

Assay time

55m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human IgG4 ELISA Kit, Chemiluminescent (ab325733) is a rapid single-wash sandwich 55-min ELISA to measure Human IgG4 in Serum, Plasma – Citrate, Plasma – EDTA, Plasma – Heparin, Cell Culture Media, Urine, Saliva, Milk with a sensitivity of 2.83 pg/mL.

How the assay works

Human IgG4 ELISA Kit, Chemiluminescent (ab325733) SimpleStep Ignite™ ELISA 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 during a 30 min only incubation, significantly reducing assay time. By utilizing a chemiluminscent detection method this improves the sensitivity and dynamic range when compared to traditional colorimetric ELISAs.

Assay Specificity

Our SimpleStep Ignite™ ELISA kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human IgG4 (ab325733) protocol summary

1. Mix : add samples / standards to the wells together with the capture and detector antibody cocktail. Incubate 30 min at room temperature
2. Wash
3. Add ChemiHRP development solution - incubate for 2 min
4. Read the results on a plate reader capable of luminescence detection

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

- Anti-IgG4 antibody [EPR26764-64] - BSA and Aside free (Capture) ab309230
- Anti-IgG4 antibody [EPR26764-24] - BSA and Aside free (Detector) ab309231
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Precision

[ { "reproducibilityType": "Intra", "sample": "Serum", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 5.4" }, { "reproducibilityType": "Inter", "sample": "Serum", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 9.4" } ]
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Recovery

[ { "sample": "Serum", "range": "81 - 91 %", "average": "= 86" }, { "sample": "Citrate plasma", "range": "103 - 114 %", "average": "= 108" }, { "sample": "EDTA Plasma", "range": "98 - 106 %", "average": "= 102" }, { "sample": "Heparin Plasma", "range": "114 - 115 %", "average": "= 114" }, { "sample": "Urine", "range": "87 - 105 %", "average": "= 94" }, { "sample": "Saliva", "range": "99 - 121 %", "average": "= 108" }, { "sample": "Milk", "range": "84 - 104 %", "average": "= 95" }, { "sample": "Cell culture media", "range": "89 - 90 %", "average": "= 90" } ]
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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Product protocols

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Target data

Constant region of immunoglobulin (Ig) heavy chains. Igs are membrane-bound or secreted glycoproteins produced by B lymphocytes. In the recognition phase of humoral immunity, the membrane-bound Igs serve as receptors, which upon binding to a specific antigen trigger the clonal expansion and differentiation of B lymphocytes into Ig-secreting plasma cells. Secreted Igs known as antibodies mediate the effector phase of humoral immunity by blocking the interaction of infectious antigens with cellular receptors (via the antigen-binding region) and eliciting effector mechanisms that lead to pathogen neutralization (via the constant region) (PubMed : 17576170, PubMed : 20176268, PubMed : 22158414). The antigen-binding region is formed by the variable domain of one heavy chain paired with the variable domain of its associated light chain. Each Ig molecule has two antigen-binding sites with remarkable affinity for a particular antigen due to V-(D)-J rearrangement, somatic hypermutations and affinity maturation of the variable domains upon antigen exposure (PubMed : 17576170, PubMed : 20176268, PubMed : 22158414). The constant region defines the Ig isotype that perform distinct sets of effector functions. B cells diversify and rearrange their Ig constant regions through class-switch recombination, a process by which the constant region is switched from one Ig isotype to another, namely from IgM and IgD to IgG, IgA and IgE (PubMed : 17576170, PubMed : 20176268, PubMed : 22158414). The constant region interacts (via the fragment crystallizable, Fc) with the Fc receptors on innate immune cells to mediate humoral effector functions. Ig gamma-4 (IgG4) isotype does not elicit antibody-dependent cellular cytotoxicity (ADDC) or complement-dependent cytotoxicity (CDC). Instead it is likely involved in immune tolerance mechanisms to allergens and parasites either by blocking IgE-antigen complex formation or by directly inhibiting mast cell degranulation through Fc receptor signaling. In the context of tumorigenesis, it may participate in immunosuppressive mechanisms.
See full target information IGHG4
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websiteProtocolBooklet
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Product promise

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