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Human IL-4 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
(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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Product Specifications
Analytical sensitivity
<0.01 pg/mL
Assay range
0.0128 - 5000 pg/mL
Sample type/volume
Serum
2-5 µL
Supernatant
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
Antibody-Conjugate A
Antibody-Conjugate B
Antibody-Conjugate Dilution Buffer
Standard
Master Mix
Ligase
Shipping conditions
Dry ice
Storage
-20°C
Protein name
IL-4
Species (tested)
Human
Assay kit format
Proximity-based Immunoassay
Label or dye
FAM™
About This Kit
The ProQuantum Human IL-4 Immunoassay Kit is designed to provide highly sensitive quantitative measurements of human IL-4 protein in small sample volumes. Utilizing proximity ligation assay (PLA) 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 Cloud-based 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 below).
The assay workflow is fast and easy-2 steps in 2 hours. There are a total of 7 components in each kit (figure below). 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 cloud-based software 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, set up customized assay instructions, and obtain robust statistical group-wise comparisons.
Target information
Interleukin-4 (IL-4) is a pleiotropic, immune-modulatory cytokine produced by activated T cells, and serves as a ligand for interleukin 4 receptor. IL-4 has been found to have the broadest range of actions. The interleukin 4 receptor also binds to IL13, which may contribute to many overlapping functions of this cytokine and IL13. STAT6, a signal transducer and activator of transcription, has been shown to play an important role in mediating the immune regulatory signal of IL-4. The IL-4 gene, IL-3, IL-5, IL-13, and CSF2 form a cytokine gene cluster on chromosome 5q. In particular, IL-4 is located in close proximity to IL13 on the chromosome. Further, the IL-4 gene, IL13 and IL5 are found to be coordinated by several long-range regulatory elements over a 120 kilobase range on the chromosome. IL-4 delta 2 is a known splice variant of IL-4.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Gene aliases : BCGF-1, BCGF1, BSF-1, BSF1, IL-4, IL4
Gene ID : (Human) 3565
Gene symbol : IL4
Protein Aliases : B cell growth factor 1, B-cell stimulatory factor 1, B_cell stimulatory factor 1, Binetrakin, BSF-1, IL-4, interleukin 4 variant 2, Interleukin-4, Lymphocyte stimulatory factor 1, Pitrakinra
UniProt ID (Human) P05112
Performance Guarantee
If an Invitrogen™ antibody 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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