Technical guide & protocol Tech Support

Product Specifications

Analytical sensitivity

<0.3 pg/mL

Assay range

0.32-5,000 pg/mL

Sample type/volume

Serum
2-5 µL
Plasma
2-5 µL
Supernatant
2-5 µL

Hands-on time

30 min

Time-to-result

2 hr

Homogenous (no wash)

Yes

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

IFN beta

Species (tested)

Mouse

Assay kit format

Proximity-based Immunoassay

Label or dye

FAM™

About This Kit

The ProQuantum Mouse IFN beta Immunoassay Kit is designed to provide quantitative measurements of mouse IFN beta 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

Type I Interferons (IFN-alpha/beta) are produced primarily in response to viral infection by Natural IFN-producing cells (IPCs) as part of the host immune response. IFNs can also inhibit the development of tumors. IFN-beta binding results in the activation of the tyrosine kinases Jak1 and Tyk2, phosphorylation of members of the STAT family of transcription factors, and the transcription and expression of the immune response genes. More recently, several members of the toll-like receptor (TLR) family were found to stimulate the production IFN-beta. IFN-beta is currently used clinically for treatment of tumors, infections and multiple sclerosis.

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

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Bioinformatics

Gene aliases : Ifb, IFN-beta, IFNB, Ifnb1

Gene ID : (Mouse) 15977

Gene symbol : Ifnb1

Protein Aliases : IFN-beta, Interferon beta

UniProt ID (Mouse) P01575

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