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IL-2Rβγ Bioassay

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Assess the Mechanism of Action of Cytokine Biologics Designed to Target the Intermediate Affinity IL-2Rβγ Receptor

  • Lacks CD25 expression
  • Prequalified according to ICH guidelines
  • 96- and 384-well plate formats
  • No primary cell culture required
  • Supplied in convenient thaw-and-use format; also available in cell propagation model (CPM) format

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IL-2Rβγ Bioassay
Thaw and Use/1X each component
$ 870.00
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Convenient IL-2Rβγ Bioassay, Consistent Results

The IL-2 receptor (IL-2R) is a type 1 cytokine receptor consisting of three subunits: IL-2Rα (CD25), IL-2/IL-15Rβ (CD122) and IL-2Rγ (CD132). The high-affinity trimeric receptor form consists of all three subunits and is found on regulatory T cells (Treg), activated CD4 and CD8 effector T cells and some natural killer (NK) cells, as well as various endothelial cell types. A medium-affinity heterodimeric receptor form, containing only IL-2Rβ and IL-2Rβγ, is found on CD8+ effector memory T cells and most NK cells. IL-2Rγ is also known as the γ common chain (γc) and is found in receptors for IL-4, IL-7, IL-9, IL-15 and IL-21.

High-dose recombinant IL-2 therapy can promote the expansion and development of immunosuppressive Treg cells, which are generally regarded as undesirable in the tumor microenvironment. IL-15 is a functionally related cytokine; however, unlike IL-2, it has no effect on Treg cells. To avoid the pitfalls of high-dose IL-2, other IL-2 and IL-15 molecules are in development that target the medium-affinity receptor and cell types that express it, which may produce a safer and more specific therapeutic response.

The IL-2Rβγ Bioassay is a bioluminescent cell-based assay designed to measure human IL-2Rβγ receptor stimulation or inhibition. The assay overcomes many of the limitations of current methods that cannot differentiate between signaling through high- and medium-affinity IL-2 receptors. These cells can be used for the development of biologic therapies intended to preferentially target the intermediate affinity heterodimeric receptor.

Bioassay Components and Product Formats

The IL-2Rβγ 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.# JA5101, JA5105).

The IL-2Rβγ Bioassay is also available in a Cell Propagation Model (CPM) (Cat.# J3392) and Cell Bank format (Cat.# GA1410) as cryopreserved cells that can be thawed and propagated for long-term use.

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

The IL-2Rβγ Bioassay is a bioluminescent cell-based assay designed to measure human IL-2Rβγ receptor stimulation or inhibition.

How the IL-2Rβγ Bioassay works.

Representation of the IL-2Rβγ Bioassay. The IL-2Rβγ Bioassay consists of a genetically engineered cell line, IL-2Rβγ Bioassay Cells. When IL-2 or IL-15 bind to the medium-affinity CD122/CD132 heterodimeric receptor complex (IL-2Rβγ), receptor-mediated pathway signaling induces luminescence that can be detected upon addition of Bio-Glo-NL™ Reagent and quantified with a luminometer. In the absence of cytokine, no signaling occurs downstream of the CD122/CD132 complex and a luminescent signal is not generated.

Assay Performance Data

Accuracy and Precision are critical to bioassays and the development of biologics. The IL-2Rβγ 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.

32454752-data-target-combined

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

Parameter Results
Accuracy
% Expected Relative Potency
% Recovery
50
90.3
75
102.6
125
105.7
150
101.7
Repeatability (% CV)
100% (Reference)
3.6
Intermediate Precision (% CV)  
8.4
Linearity (r2)  
0.996
Linearity (y = mx + b)  
y = 1.08x – 6.35
A 50–150% theoretical potency series of recombinant human IL-15 was analyzed in triplicate in three independent experiments performed on three days by two analysts (for a total of six independent experiments). Bio-Glo-NL™ 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.
Representative data for the IL-2Rβγ Bioassay; EC50 = 81ng/ml.
18834ma-c

Representative data for the IL-2Rβγ Bioassay. IL-2Rβγ Bioassay cells were prepared as described in the protocol and incubated with serial dilutions of recombinant IL-2 and IL-15. After a 6-hour incubation, Bio-Glo-NL™ Reagent was added, and luminescence was quantified using the GloMax® Discover System. Data were fitted to a 4PL curve using GraphPad Prism® software. The IL-2 EC50 was 81ng/ml (Panel A), with a fold induction of approximately 24 (Panel B). The IL-15 EC50 was 4.4ng/ml with a fold induction of approximately 28. Data were generated using cells from culture. Panel A shows raw luminescence measurements. Panel B displays the calculated fold induction.

Specifications

You are viewing: JA5101 Change Configuration

Whats in the box

Item Part # Size

IL-2Rβγ Bioassay Cells

J340A 1 × 0.6ml

Fetal Bovine Serum

J121A 1 × 4ml

RPMI 1640 Medium

G708A 1 × 36ml

Bio-Glo-NL™ Luciferase Assay Buffer

J308A 1 × 10ml

Bio-Glo-NL™ Luciferase Assay Substrate

J309A 1 × 200μl

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

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-NL™ 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. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.

Product cannot be used for proficiency testing.

Specifications

You are viewing: JA5105 Change Configuration

Whats in the box

Item Part # Size

IL-2Rβγ Bioassay Cells

J340A 5 × 0.6ml

Fetal Bovine Serum

J121A 5 × 4ml

RPMI 1640 Medium

G708A 5 × 36ml

Bio-Glo-NL™ Luciferase Assay Buffer

J308A 5 × 10ml

Bio-Glo-NL™ Luciferase Assay Substrate

J309A 5 × 200μl

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

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-NL™ 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. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.

Product cannot be used for proficiency testing.

Specifications

You are viewing: J3392 Change Configuration

Whats in the box

Item Part # Size

IL-2Rβγ Bioassay Cells (CPM)

J339A 2 × 1ml

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

XX

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-NL™ 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. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.

Product cannot be used for proficiency testing.

Specifications

You are viewing: GA1410 Change Configuration

Whats in the box

Item Part # Size

IL-2Rβγ Bioassay Cells (CPM)

J339A 50 × 1ml

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

XX

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-NL™ 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. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.

Product cannot be used for proficiency testing.

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