IL-2 Bioassay
A Simpler, More Consistent IL-2 Bioassay
- Prequalified according to ICH guidelines
- Mechanism-of-action-based assay
- No primary cell culture required
- Supplied in convenient thaw-and-use format; also available in cell propagation model (CPM) format
Catalog Number:
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Catalog Number: JA2201
Catalog Number: JA2205
Catalog Number: J2952
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
'Cells as reagents' Method
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
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.
IL-2 Bioassay Cells were incubated with serial dilutions of recombinant IL-2. After a 6-hour incubation, Bio-Glo™ Reagent was added, and luminescence was quantified using the GloMax® Discover System.
Proleukin® (aldesleukin; Clinigen, 1.1mg/ml undiluted) was heat stressed at 53°C prior to being tested in the IL-2 Bioassay. Bio-Glo™ Reagent was added, and luminescence was quantified using the GloMax® Discover System.
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).
Specifications
Catalog Number:
What's in the box
| Item | Part # | Choose a 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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Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
Please see the Protocol for detailed storage conditions.
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.
What's in the box
| Item | Part # | Choose a 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 SDSCertificate of Analysis
Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
Please see the Protocol for detailed storage conditions.
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.
What's in the box
| Item | Part # | Choose a size |
|---|---|---|
IL-2 Bioassay Cells (CPM) |
J295A | 2 × 0.75ml |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
Please see the Protocol for detailed storage conditions.
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.
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Frequently Used With
Bio-Glo™ Luciferase Assay System
Bioluminescent detection reagent qualified for use with bioassays that utilize firefly luciferase.
G7940, G7941
GloMax® Discover System
High-performance microplate reader for detecting luminescence, fluorescence and absorbance.
GM3000
RealTime-Glo™ MT Cell Viability Assay
A bioluminescent method to kinetically monitor viability in cell culture up to 72 hours.
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