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RANKL Bioassay

Measure Potency and Stability of Biologics Designed to Affect RANKL Signaling

  • 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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RANKL Bioassay
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Simple RANKL Bioassay, Consistent Results

Receptor activator of nuclear factor-κB (RANK/TRANCE receptor/TNFRSF11A) is a member of the tumor necrosis factor receptor (TNFR) family. Binding of its ligand RANKL to the receptor regulates osteoclast formation, activation and survival in bone modeling and remodeling, along with several pathologic conditions characterized by increased bone turnover.

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

Assay Components

RANKL 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.# JA2701, JA2705).

RANKL 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.# J3102).

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

Schematic of the RANKL Bioassay. 16461ma-01

Representation of the RANKL Bioassay. The RANKL Bioassay consists of a genetically engineered cell line, RANKL Bioassay Cells. When RANKL binds to its receptor, receptor-mediated pathway signaling induces luminescence that can be detected upon the addition of Bio-Glo™ Reagent and quantified with a luminometer. Inhibition of RANKL binding by anti-RANKL antibody 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 RANKL Bioassay Shows Precision, Accuracy and Linearity.

Parameter Results
Accuracy % Expected Relative Potency % Recovery
50 54.1
75 79.9
125 128.4
150 152.7
Repeatability (% CV) 100% (Reference) 1.79
Intermediate Precision (% CV)   5.98
Linearity (r2)   0.999
Linearity (y = mx + b)   y = 0.9819x – 5.5733
A 50–150% theoretical potency series of denosumab (anti-RANKL) 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 was 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.

 

Data graph showing that the RANKL Bioassay responds to recombinant RANKL.
The RANKL Bioassay responds to recombinant RANKL. RANKL Bioassay Cells were grown and prepared as described in this protocol, and incubated with serial dilutions of recombinant RANKL. After a 6-hour incubation, Bio-Glo™ Reagent was added, and luminescence was quantified using the GloMax® Discover System. This graph shows raw luminescence measurements. 
Data graph showing that RANKL Bioassay responds to denosumab.
The RANKL Bioassay responds to denosumab. RANKL Bioassay Cells were grown and prepared as described in this protocol, and incubated with serial dilutions of denosumab (anti-RANKL) antibody and 100ng/ml RANKL. After a 6-hour incubation, Bio-Glo™ Reagent was added, and luminescence was quantified using the GloMax® Discover System. Data were generated using thaw-and-use cells.
  • Kits are available in 1X and 5X sizes.

Specifications

You are viewing: J3102 Change Configuration

What's in the box?

Item Part # Size

RANKL Bioassay Cells (CPM)

J310A 2 × 1ml

SDS

Choose language:

Download SDSPDF (195 KB) – English (United States)

Certificate of Analysis

Search by lot number

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.

Patent Pending.

Product cannot be used for proficiency testing.

Specifications

You are viewing: JA2701 Change Configuration

What's in the box?

Item Part # Size

RANKL Bioassay Cells

J312A 1 × 0.5ml

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 (303 KB) – English (United States)

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 and European Pat. No. 1341808.

Patent Pending.

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

Licensed from Lonza Cologne GmbH under U.S. Pat. Nos. 7,700,357, 8,192,990 and 8,003,389, European Pat. Nos. 1297119, 1522587, 1607484 and 1741778 and other pending and issued patents.

Product cannot be used for proficiency testing.

Specifications

You are viewing: JA2705 Change Configuration

What's in the box?

Item Part # Size

RANKL Bioassay Cells

J312A 5 × 0.5ml

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 (304 KB) – English (United States)

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. 10,077,244 and other patents.

Patent Pending.

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

Product cannot be used for proficiency testing.

Licensed from Lonza Cologne GmbH under U.S. Pat. Nos. 7,700,357, 8,192,990 and 8,003,389, European Pat. Nos. 1297119, 1522587, 1607484 and 1741778 and other pending and issued patents.

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