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IL-6 Bioassay

A Simpler, More Consistent IL-6 Bioassay

  • 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-6 Bioassay
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Measure the Ability of Biologics to Activate or Inhibit Cytokine Signals

IL-6 is a member of the IL-6 cytokine family, which includes IL-11, leukemia inhibitory factor (LIF), oncostatin M, ciliary neurotrophic factor (CNTF), cardiotrophin-1 (CT-1) and cardiotrophin-like cytokine (CLC). All of these cytokines signal through gp130 and the STAT3 pathway. IL-6 is transiently secreted following tissue damage or stressors, including UV irradiation, reactive oxygen species, and microbial and viral agents. It is also one of the cytokines released during bacterial sepsis. Furthermore, IL-6, along with transforming growth factor β (TGFβ), promotes differentiation of CD4+ T cells into Th17 cells and inhibits differentiation of regulatory T cells, thus playing a critical role in autoimmunity. 

The IL-6 Bioassay is a bioluminescent cell-based assay designed to measure IL-6 stimulation or inhibition. The IL-6 Bioassay Cells have been engineered to express luc2P in response to IL-6 signaling. When IL-6 binds to IL-6 Bioassay Cells, the receptor transduces intracellular signals, resulting in luminescence. The bioluminescent signal is detected and quantified using Bio-Glo™ Reagent.

Assay Components

The IL-6 Bioassay Cells 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.# JA2501, JA2505).

IL-6 Bioassay Cells are also available in a Cell Propagation Model (CPM) format, as cryopreserved cells that can be thawed, propagated and banked for long-term use (Cat.# J2992).

Workflow

Standard bioassay workflow with cultured cells
Improved bioassay workflow with thaw-and-use cells

Standardized Reagents Improve Results

  • Thaw-and-use 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.

Assay Principle

IL-6 Bioassay schematic diagram.

Representation of the IL-6 Bioassay. The IL-6 Bioassay consists of a genetically engineered cell line, IL-6 Bioassay Cells. When IL-6 binds to the IL-6 receptor (IL-6R), receptor-mediated pathway signaling induces luminescence that can be detected upon addition of Bio-Glo™ Reagent, and quantified with a luminometer. Inhibition of IL-6 binding by anti-IL-6 or anti-IL-6R antibodies results in a decrease in luminescence.

Assay Performance Data

Accuracy and precision are critical to bioassays and the development of biologics. 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 how close results are to 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 the assay to provide measured values that are proportional to the concentration of the sample.

32454752-data-target-combined

The IL-6 Bioassay Shows Precision, Accuracy and Linearity.

Parameter Results
Accuracy % Expected Relative Potency % Recovery
50 104
75 101.3
125 107
150 106
Repeatability (% CV) 100% (Reference) 2.93
Intermediate Precision (% CV)   11.8
Linearity (r2)   0.999
Linearity (y = mx + b)   y = 1.087x – 3.54
A 50–150% theoretical potency series of tocilizumab (anti-IL-6R) was analyzed in triplicate in three independent experiments performed on three days by two analysts. Bio-Glo™ Reagent was added and luminescence quantified using the GloMax® Discover Detection System. Data were analyzed and relative potencies calculated after parallelism determination using JMP® software. Data were generated using thaw-and-use cells.

 

The IL-6 Bioassay responds to recombinant IL-6.
The IL-6 Bioassay responds to recombinant IL-6.

IL-6 Bioassay Cells were grown and prepared as described in the protocol, and incubated with serial dilutions of recombinant IL-6. After a 4-, 5-, 6- or 24-hour incubation, Bio-Glo™ Reagent was added, and luminescence was quantified using the GloMax® Discover System.

The IL-6 Bioassay indicates stability.
The IL-6 Bioassay indicates stability.

Tocilizumab (anti-IL-6R) was treated with various concentrations of hydrogen peroxide (0–0.05%) for 18 hours at 26°C prior to use in the IL-6 Bioassay. Bio-Glo™ Reagent was added, and luminescence was quantified using the GloMax® Discover System.

Product Kit Formats

  • Kits are available in 1X and 5X sizes.

Specifications

You are viewing: J2992 Change Configuration

What's in the box?

Item Part # Size

IL-6 Bioassay Cells (CPM)

J299A 2 × 0.65ml

SDS

Choose language:

Download SDSPDF (199 KB) – English (United States)

Use Restrictions

Not For Medical Diagnostic Use.

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™ 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 and European Pat. No. 1341808.

Patent Pending.

Cells were obtained under license from AdVec Inc.

Specifications

You are viewing: JA2501 Change Configuration

What's in the box?

Item Part # Size

IL-6 Bioassay Cells

J302A 1 × 0.65ml

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

Choose language:

Download SDSPDF (301 KB) – English (United States)

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 and European Pat. No. 1341808.

Patent Pending.

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

Cells were obtained under license from AdVec Inc.

Specifications

You are viewing: JA2505 Change Configuration

What's in the box?

Item Part # Size

IL-6 Bioassay Cells

J302A 5 × 0.65ml

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

Choose language:

Download SDSPDF (302 KB) – English (United States)

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. 10,077,244 and other patents.

Patent Pending.

U.S. Pat. No. 8,008,006 and European Pat. No. 1341808.

Cells were obtained under license from AdVec Inc.

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