IL-2 Bioassay

Simple IL-2 Bioassay, Consistent Results

Interleukin-2 (IL-2), originally described as “T-cell growth factor” in 1976, is a small 15.5kDa monomer secreted by a variety of cell types including CD4+ and CD8+ T cells, natural killer (NK) cells and activated dendritic cells. IL-2 has pleiotropic effects on the immune system. It plays a critical role in the generation, maintenance and expansion of CD4+ regulatory T cells, promotes the cytotoxic activity of NK and CD8+ cells and governs homeostasis through the elimination of harmful autoreactive T cells via activation-induced cell death.

The IL-2 Bioassay is a bioluminescent cell-based assay designed to measure IL-2 stimulation or inhibition. The assay overcomes many of the limitations of the current endothelial cell proliferation assays and can be used for the discovery and development of novel biologic therapies aimed at either inducing or inhibiting the IL-2 response. The IL-2 Bioassay Cells have been engineered to express luc2 in response to IL-2 signaling. When IL-2 binds to IL-2 Bioassay Cells, the receptor transduces intracellular signals that result in luminescence. The bioluminescent signal is detected and quantified using Bio-Glo™ Luciferase Assay System (Cat.# G7940) and a standard luminometer such as the GloMax® Discover System (Cat.# GM3000).

Assay Components

The IL-2 Bioassay Cells included are provided in a thaw-and-use format as cryopreserved cells that can be thawed, plated and used in an assay without the need for cell propagation (Cat.# JA2201, JA2205).

The IL-2 Bioassay is also available in a Cell Propagation Model (CPM) format, as cryopreserved cells, that can be thawed, propagated and banked for long-term use (Cat.# J2952).

The Workflow

Traditional Method

Standard bioassay workflow with cultured cells

'Cells as reagents' Method

Improved bioassay workflow with thaw-and-use cells

Standardized Reagents Improve Results

Thaw-and-use cell format provides significant time and labor savings over traditional methods while reducing variability.
Kit includes all of the required reagents in standardized formats.
Easily amenable to high-throughput workflows.

The Assay Principle

How the IL-2 Bioassay works. 15414ma — **Representation of the IL-2 Bioassay.** The IL-2 Bioassay consists of a genetically engineered cell line, IL-2 Bioassay Cells. When IL-2 binds to its receptor, receptor-mediated pathway signaling induces luminescence that can be detected upon addition of Bio-Glo™ Reagent and quantified with a luminometer. In the absence of IL-2, no signaling occurs downstream of IL-2R and no luminescent signal is generated.

Assay Performance Data

Accuracy and Precision are critical to bioassays and the development of biologics. The IL-2 Bioassay is both accurate and precise. Accuracy is a measure of how close results are to a target or reference value. Precision is a measure of repeatability. This assay was developed and prequalified according to ICH guidelines for accuracy and precision, giving it the performance characteristics required for many applications of a bioassay used for antibody drug discovery research and development.

Accuracy: Measure of ability to replicate a reference or "target" value.

IntraLab Precision: Measure of repeatability from multiple assays of the same homogeneous sample under the same operating conditions over a short time (same analyst; same lab; same day).

Intermediate Precision: Measure of repeatability of multiple assays of the same homogeneous sample with different analysts or equipment or on different days.

Linearity: Ability of your assay to provide measured values that are proportional to the concentration of the sample.

The IL-2 Bioassay Shows Precision, Accuracy and Linearity.

Parameter	Results
Accuracy	% Expected Relative Potency	% Recovery
	50	103.9
	70	99.8
	140	100.5
	200	101.6
Repeatability (% CV)	100% (Reference)	4.32
Intermediate Precision (% CV)		6.88
Linearity (r²)		0.999
Linearity (y = mx + b)		y = 1.01x + 0.09
A 50–200% theoretical potency series of aldesleukin (therapeutic IL-2) was analyzed in triplicate in three independent experiments performed on three days by two analysts (for a total of six independent experiments). Bio-Glo™ Reagent was added and luminescence quantified using the GloMax® Discover System. Data were analyzed and relative potencies calculated after parallelism determination using JMP® software. Data were generated using thaw-and-use cells.

IL-2 Bioassay is responsive to recombinant IL-2.

The IL-2 Bioassay indicates stability.

Product Kit Formats and Related Products

IL-2 Bioassay is available in 1X and 5X sizes.
IL-2 Bioassay is also available in CPM format (Cat.# J2952).

Protocols

Complete Protocol

IL-2 BioassayPDF (1740 KB) – English

IL-2 Bioassay, Propagation ModelPDF (1172 KB) – English

What's in the box?

Item	Part #	Size
IL-2 Bioassay Cells	J297A	1 × 0.80ml
Fetal Bovine Serum	J121A	1 × 4ml
RPMI 1640 Medium	G708A	1 × 36ml
Bio-Glo™ Luciferase Assay Buffer	G719A	1 × 10ml
Bio-Glo™ Luciferase Assay Substrate	G720A	1 × 1 vial

SDS

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Certificate of Analysis

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Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.

This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use” means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to propagate, modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega’s. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

U.S. Pat. No. 8,008,006.

Patent Pending.

U.S. Pat. No. 10,077,244 and other patents.

Product cannot be used for proficiency testing.

What's in the box?

Item	Part #	Size
IL-2 Bioassay Cells	J297A	5 × 0.80ml
Fetal Bovine Serum	J121A	5 × 4ml
RPMI 1640 Medium	G708A	5 × 36ml
Bio-Glo™ Luciferase Assay Buffer	G719A	5 × 10ml
Bio-Glo™ Luciferase Assay Substrate	G720A	5 × 1 vial

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.

This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use” means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to propagate, modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega’s. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

U.S. Pat. No. 8,008,006.

Patent Pending.

U.S. Pat. No. 10,077,244 and other patents.

Product cannot be used for proficiency testing.

What's in the box?

Item	Part #	Size
IL-2 Bioassay Cells (CPM)	J295A	2 × 0.75ml

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

Please see the Protocol for detailed storage conditions.

Patents and Disclaimers

NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the recipient is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the recipient with a full refund.

This product may not be further sold or transferred by the recipient and may be used only by the recipient, and then only for (1) research use, (2) discovery, development and monitoring of biologic drugs and vaccines, (3) quality assurance testing of biologic drugs and vaccines, and (4) product release assays for biologic drugs and vaccines. No other commercial use is allowed. "Commercial use" means any and all uses of this product by recipient for monetary or other consideration, including providing a service, information or data to unaffiliated third parties, and resale of this product for any use. Recipient has no right to modify, derivatize, genetically engineer or otherwise create variations of the cells or genes stably transfected within the cells except that recipient may propagate and store the cells for long-term use. In addition, recipient must use Bio-Glo™ Luciferase Assay System purchased from Promega Corporation for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega's. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING AS TO MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THIS PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

U.S. Pat. No. 8,008,006.

Product cannot be used for proficiency testing.

Resources

Posters

Other Resources

Tips and Tools for Successful Immunotherapy Bioassay Development

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IL-2 Bioassay

A Simpler, More Consistent IL-2 Bioassay

Simple IL-2 Bioassay, Consistent Results

Assay Components

The Workflow

The Assay Principle

Assay Performance Data

IL-2 Bioassay is responsive to recombinant IL-2.

The IL-2 Bioassay indicates stability.

Product Kit Formats and Related Products

Protocols

Complete Protocol

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Patents and Disclaimers

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Patents and Disclaimers

Specifications

What's in the box?

SDS

Certificate of Analysis

Use Restrictions

Storage Conditions

Patents and Disclaimers

Resources

Posters

Other Resources

Similar Products

IL-15 Bioassay

VEGF Bioassay

Frequently Used With

Bio-Glo™ Luciferase Assay System

GloMax® Discover System

RealTime-Glo™ MT Cell Viability Assay

CellTiter-Glo® 2.0 Cell Viability Assay

pHAb Reactive Dyes

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