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Human IgG4 ProQuantum Immunoassay Kit

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Technical guide & protocol Tech Support
Human IgG4 ProQuantum Immunoassay Kit
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Human IgG4 ProQuantum Immunoassay Kit
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Human IgG4 ProQuantum Immunoassay Kit

How ProQuantum immunoasssays work
(A) Antibody-antigen binding: Antibodies bind to two separate epitopes on the antigen (during a 1-hr incubation), which brings the two conjugated oligonucleotides into close proximity. (B) Ligation and amplification of signal (in a qPCR instrument): DNA ligase and a third splint oligonucleotide connector are added to ligate the two ends of the conjugated oligonucleotides, creating a 100-base DNA template. Once the ligase is inactivated at 95°C, the sample is amplified through 40 cycles of annea... View More
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Human IgG4 ProQuantum Immunoassay Kit
Human IgG4 ProQuantum Immunoassay Kit
Human IgG4 ProQuantum Immunoassay Kit (A45247)
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Product Specifications

Analytical sensitivity

<0.05 ng/mL

Assay range

0.08-1250 ng/mL

Sample type/volume

Serum
2-5 µL
Plasma
2-5 µL

Hands-on time

30 min

Time-to-result

2 hr

Homogenous (no wash)

Yes

Instrument

qPCR Instruments

Product size

96 Tests

Contents

Assay Dilution Buffer
Antibody-Conjugate A
Antibody-Conjugate B
Antibody-Conjugate Dilution Buffer
Standard
Master Mix
Ligase

Shipping conditions

Dry ice

Storage

-20°C

Protein name

Human IgG4

Species (tested)

Human

Assay kit format

Proximity-based Immunoassay

Label or dye

FAM™

About This Kit

The ProQuantum Human IgG4 Immunoassay Kit is designed to provide quantitative measurements of human IgG4 in small sample volumes. Utilizing proximity-based amplification technology, the assay combines the analyte specificity of high-affinity antibody-antigen binding with the signal detection and amplification capabilities of real-time PCR to achieve a simple yet powerful next-generation protein quantitation platform. A user-friendly workflow combined with intuitive software for analytics enables sample-to-answer in just 2 hours.

• High sensitivity-detect low levels of protein with greater sensitivity than traditional methods

• Broad dynamic range-5 logarithmic units, minimizing sample dilutions to ensure they fall within the range

• Small sample consumption-use 2-5 µL of sample (compared to 75 µL for triplicate wells with other methods)

• Fast, easy workflow-2 hours from sample to answer, with no wash steps

• No proprietary instrument to purchase-runs on any real-time PCR instrument

ProQuantum immunoassays utilize a matched pair of target-specific antibodies, each conjugated to a DNA oligonucleotide. During antibody-analyte binding, the two DNA oligos are brought into close proximity, which allows for ligation of the two strands and subsequent creation of a template strand for amplification. This platform leverages the sensitivity and large dynamic range of Applied Biosystems TaqMan real-time PCR technology (Figure 1).

The assay workflow is fast and easy-2 steps in 2 hours. There are a total of 7 components in each kit (Figure 2). First, mix the antibody-conjugates, dilute the sample, and create the standard curve in a working plate. Then, using a multi-channel pipette, add the antibody-conjugates and sample (or standard) into the wells of a PCR plate and incubate for 1 hour. Combine the master mix and ligase and add to the wells of the PCR plate, then run the plate on any qPCR instrument. After the run is complete, import the results file into the ProQuantum software (cloud-based version at https://apps.thermofisher.com/apps/proquantum). Using this software, the data can be analyzed easily to obtain protein concentration values. The software allows you to set up standard curves, design plate layouts, apply a 5PL weighted algorithm, and obtain robust statistical group-wise comparisons.

Target information

Human IgG4, also known as human immunoglobulin G4, is a subclass of human IgG antibodies that is produced by the immune system in response to antigens. It is the least abundant subclass of IgG in the blood and plays a minor role in defense against pathogens. IgG4 works by binding to specific antigens and promoting their clearance from the body through various mechanisms, including phagocytosis and complement activation. IgG4 is also involved in other immune responses, such as immune tolerance and suppression of inflammation. Human IgG4 can be used in research to study the immune response to pathogens and to develop diagnostic tests and therapeutic interventions for infectious and autoimmune diseases.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Gene ID : (Human) 3503

Gene symbol : IGHG4

Protein Aliases : ImmunoglobulinG, Immunoglobulin G, Immunoglobulin G4

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SIZE

10 x 96 tests

PRICE

USD 400.00

Cat # CHC1263

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Performance Guarantee

If an Invitrogen™ immunoassay doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*

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