TLR3 Bioassay
Measure Cellular TLR3 Signaling from Double-Stranded RNA Contamination in IVT mRNA Samples
- Specific: No interference from other TLRs
- Differentiable: Response dependent on dsRNA size
- Simple: Add-mix-read protocol
- Convenient: Reagents provided in Thaw-and-Use format
Catalog Number:
Choose a Product Format
Size
Catalog Number: CS355505
Catalog Number: CS355507
Catalog Number: CS355502
TLR3 Bioassay: Streamlined dsRNA Detection for Quality Assurance
The presence of double-stranded RNA (dsRNA) in in vitro transcription (IVT) products is a significant challenge for the development of RNA-based therapeutics. As a common byproduct of IVT, dsRNA contamination can trigger downstream immunogenic responses, which therapeutic developers seek to minimize. Measuring and controlling dsRNA levels is critical to optimizing mRNA vaccine and therapeutic production, ensuring the safety and efficacy of these life-saving technologies.
The TLR3 Bioassay offers a cutting-edge solution for precise, MoA-based detection of dsRNA contaminants. This easy-to-use assay is available in a thaw-and-use format, which eliminates the need for cell culture, providing rapid and reliable results. Its ability to detect variations in dsRNA size, sequence and delivery methods make it an invaluable tool for ensuring the quality of mRNA preparations, enabling researchers to streamline workflows and improve manufacturing outcomes.
Accurate dsRNA Contaminant Detection Made Simple
Principle of TLR3 Bioassay. TLR3 reporter cells uptake dsRNA through endocytosis. The binding of the dsRNA to endosomal TLR3 leads to the activation of the TLR3-responsive promoter, which drives luciferase reporter expression and luminescent output.
Assay Workflow: Standardized Reagents to Improve Your Results
- Thaw-and-use cell format provides significant time and labor savings over traditional methods while reducing variability.
- User-friendly protocol for quicker enablement.
- Easily amenable to high-throughput workflows.
TLR3 Bioassay Response is Specific to TLR3 and dsRNA
TLR3 Bioassay is specific to TLR3 signaling and dsRNA detection. Panel A. The TLR3 Bioassay was performed with Lipofectamine™ RNAiMAX-complexed Poly(I:C) (LMW or HMW, as indicated). Parental reporter cells that lack TLR3 expression were used as control. Panel B. The TLR3 Bioassay was performed with Poly(I:C) or Poly(U) with or without the lipid transfection reagent (Lipofectamine™ RNAiMAX). After incubation, the Bio-Glo-NL™ Reagent was added and luminescence determined in a GloMax® Discover Microplate Reader.
TLR3 Bioassay Reveals Differential Immune Activation by dsRNA Size and Delivery Method
TLR3 Bioassay is suitable for comparing immune responses from different species of dsRNA contaminants. The TLR3 Bioassay was performed with (Panel A) Lipofectamine™ RNAiMAX-complexed dsRNA samples of different size and sequence or (Panel B) Poly(I:C) (0.2–1kb) with different lipid transfection reagents. After incubation, the Bio-Glo-NL™ Reagent was added and luminescence determined in a GloMax® Discover Microplate Reader.
TLR3 Bioassay Detects dsRNA Contaminants in IVT mRNA Samples
The TLR3 Bioassay was performed with two IVT products as samples complexed with Lipofectamine™ RNAiMAX.
IVT 1: purified with LiCl2 (0.4kb product)
IVT 2: DEAE column-purified (1.2kb)
Protocols
No protocols available
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size |
|---|---|---|
TLR3 Bioassay Cells (T&U) |
CS355503 | 1 × 0.5ml |
Fetal Bovine Serum |
J121A | 1 × 4ml |
DMEM with 4.5g/L Glucose, L-Glutamine, Sodium Pyruvate |
J236A | 1 × 60ml |
Bio-Glo-NL™ Luciferase Assay Buffer |
J308A | 1 × 10ml |
Bio-Glo-NL™ Luciferase Assay Substrate |
J309A | 1 × 200μl |
SDS
Search for SDSCertificate of Analysis
Promega Cell Line Limited Use Label License (LULL)
BY USE OF THIS MATERIAL, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE.
-
1. Usage and Transfer Restrictions:
- 1.1. Recipient may use this material for research use only, which includes but is not limited to drug discovery and development, quality assurance testing and product release assays. No transfer, resale or commercial use of this material is allowed without the written consent of Promega Corporation.
- 1.2. “Commercial use” means any and all uses of this material or derivatives by recipient for monetary or other consideration, including product manufacture, providing a service, information or data to unaffiliated third parties and resale of the material for any use.
-
2. Modification Restrictions:
- 2.1. Recipient shall have no right to modify, derivatize or otherwise create variations of the nucleotide sequence of any proprietary gene, including but not limited to luciferase, NanoBiT® technology (e.g., HiBiT), HaloTag® technology or genes stably transfected within the cells.
- 2.2. Recipient shall have no right to genetically engineer or otherwise modify the cell line.
-
3. Propagation and Storage:
- 3.1. The recipient may not propagate the cells, unless material is marked as Propagation Model, Cell Bank or Master Cell Bank; then recipient may propagate and store the cells for long-term use.
-
4. For all determinations of luminescence or other assay-related activity of this material and its derivatives, recipient must either:
- 4.1. Use the detection systems specified in the protocol or data report and purchased from Promega for all luminescence assays when using this material; or
- 4.2. Contact Promega to obtain a license for use of this material with reagents other than a Bio-Glo™ branded luciferase reagent, including but not limited to:
- 4.2.1 Bio-Glo™, Bio-Glo-NL™, Bio-Glo-NB™ TCK, Bio-Glo-NB™ VLP, Bio-Glo-NB™ Lytic
-
5. For uses of this material for energy transfer (such as bioluminescence resonance energy transfer), recipient must:
- 5.1. Use NanoBRET™-branded luminescent assay reagents (LARs; e.g., NanoBRET™ Nano-Glo® Substrate), Intracellular TE Nano-Glo® Substrate/Inhibitor or Intracellular TE Nano-Glo® Vivazine™/Inhibitor for all determinations of luminescent activity by this material and its derivatives; and
- 5.2. Use NanoBRET™-branded energy acceptors (e.g., NanoBRET™ tracers, NanoBRET™ dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this material and its derivatives; or
- 5.3. Contact Promega to obtain a license for the use of the material and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded autofluorescent protein.
-
6. For uses of HaloTag® Technology in this material, recipient must either:
- 6.1. Use Promega HaloTag® ligands, which can be modified or linked to Promega or customer-supplied moieties; or
- 6.2. Contact Promega to obtain a license if Promega HaloTag® ligands are not to be used.
-
7. Disclaimer and Governing Law:
- 7.1. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE MATERIAL. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
What's in the box?
| Item | Part # | Size |
|---|---|---|
TLR3 Bioassay Cells (T&U) |
CS355503 | 5 × 0.5ml |
Fetal Bovine Serum |
J121A | 5 × 4ml |
DMEM with 4.5g/L Glucose, L-Glutamine, Sodium Pyruvate |
J236A | 5 × 60ml |
Bio-Glo-NL™ Luciferase Assay Buffer |
J308A | 5 × 10ml |
Bio-Glo-NL™ Luciferase Assay Substrate |
J309A | 5 × 200μl |
SDS
Search for SDSCertificate of Analysis
Promega Cell Line Limited Use Label License (LULL)
BY USE OF THIS MATERIAL, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE.
-
1. Usage and Transfer Restrictions:
- 1.1. Recipient may use this material for research use only, which includes but is not limited to drug discovery and development, quality assurance testing and product release assays. No transfer, resale or commercial use of this material is allowed without the written consent of Promega Corporation.
- 1.2. “Commercial use” means any and all uses of this material or derivatives by recipient for monetary or other consideration, including product manufacture, providing a service, information or data to unaffiliated third parties and resale of the material for any use.
-
2. Modification Restrictions:
- 2.1. Recipient shall have no right to modify, derivatize or otherwise create variations of the nucleotide sequence of any proprietary gene, including but not limited to luciferase, NanoBiT® technology (e.g., HiBiT), HaloTag® technology or genes stably transfected within the cells.
- 2.2. Recipient shall have no right to genetically engineer or otherwise modify the cell line.
-
3. Propagation and Storage:
- 3.1. The recipient may not propagate the cells, unless material is marked as Propagation Model, Cell Bank or Master Cell Bank; then recipient may propagate and store the cells for long-term use.
-
4. For all determinations of luminescence or other assay-related activity of this material and its derivatives, recipient must either:
- 4.1. Use the detection systems specified in the protocol or data report and purchased from Promega for all luminescence assays when using this material; or
- 4.2. Contact Promega to obtain a license for use of this material with reagents other than a Bio-Glo™ branded luciferase reagent, including but not limited to:
- 4.2.1 Bio-Glo™, Bio-Glo-NL™, Bio-Glo-NB™ TCK, Bio-Glo-NB™ VLP, Bio-Glo-NB™ Lytic
-
5. For uses of this material for energy transfer (such as bioluminescence resonance energy transfer), recipient must:
- 5.1. Use NanoBRET™-branded luminescent assay reagents (LARs; e.g., NanoBRET™ Nano-Glo® Substrate), Intracellular TE Nano-Glo® Substrate/Inhibitor or Intracellular TE Nano-Glo® Vivazine™/Inhibitor for all determinations of luminescent activity by this material and its derivatives; and
- 5.2. Use NanoBRET™-branded energy acceptors (e.g., NanoBRET™ tracers, NanoBRET™ dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this material and its derivatives; or
- 5.3. Contact Promega to obtain a license for the use of the material and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded autofluorescent protein.
-
6. For uses of HaloTag® Technology in this material, recipient must either:
- 6.1. Use Promega HaloTag® ligands, which can be modified or linked to Promega or customer-supplied moieties; or
- 6.2. Contact Promega to obtain a license if Promega HaloTag® ligands are not to be used.
-
7. Disclaimer and Governing Law:
- 7.1. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE MATERIAL. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
What's in the box?
| Item | Part # | Size |
|---|---|---|
TLR3 Bioassay Cells (CPM) |
CS355501 | 2 × 1ml |
Promega Cell Line Limited Use Label License (LULL)
BY USE OF THIS MATERIAL, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE.
-
1. Usage and Transfer Restrictions:
- 1.1. Recipient may use this material for research use only, which includes but is not limited to drug discovery and development, quality assurance testing and product release assays. No transfer, resale or commercial use of this material is allowed without the written consent of Promega Corporation.
- 1.2. “Commercial use” means any and all uses of this material or derivatives by recipient for monetary or other consideration, including product manufacture, providing a service, information or data to unaffiliated third parties and resale of the material for any use.
-
2. Modification Restrictions:
- 2.1. Recipient shall have no right to modify, derivatize or otherwise create variations of the nucleotide sequence of any proprietary gene, including but not limited to luciferase, NanoBiT® technology (e.g., HiBiT), HaloTag® technology or genes stably transfected within the cells.
- 2.2. Recipient shall have no right to genetically engineer or otherwise modify the cell line.
-
3. Propagation and Storage:
- 3.1. The recipient may not propagate the cells, unless material is marked as Propagation Model, Cell Bank or Master Cell Bank; then recipient may propagate and store the cells for long-term use.
-
4. For all determinations of luminescence or other assay-related activity of this material and its derivatives, recipient must either:
- 4.1. Use the detection systems specified in the protocol or data report and purchased from Promega for all luminescence assays when using this material; or
- 4.2. Contact Promega to obtain a license for use of this material with reagents other than a Bio-Glo™ branded luciferase reagent, including but not limited to:
- 4.2.1 Bio-Glo™, Bio-Glo-NL™, Bio-Glo-NB™ TCK, Bio-Glo-NB™ VLP, Bio-Glo-NB™ Lytic
-
5. For uses of this material for energy transfer (such as bioluminescence resonance energy transfer), recipient must:
- 5.1. Use NanoBRET™-branded luminescent assay reagents (LARs; e.g., NanoBRET™ Nano-Glo® Substrate), Intracellular TE Nano-Glo® Substrate/Inhibitor or Intracellular TE Nano-Glo® Vivazine™/Inhibitor for all determinations of luminescent activity by this material and its derivatives; and
- 5.2. Use NanoBRET™-branded energy acceptors (e.g., NanoBRET™ tracers, NanoBRET™ dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this material and its derivatives; or
- 5.3. Contact Promega to obtain a license for the use of the material and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded autofluorescent protein.
-
6. For uses of HaloTag® Technology in this material, recipient must either:
- 6.1. Use Promega HaloTag® ligands, which can be modified or linked to Promega or customer-supplied moieties; or
- 6.2. Contact Promega to obtain a license if Promega HaloTag® ligands are not to be used.
-
7. Disclaimer and Governing Law:
- 7.1. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE MATERIAL. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
Resources
Featured Resource
For over 40 years we have been a leading provider of in vitro transcription reagents supporting a variety of applications.
To better serve and support the needs of our customers in the rapidly growing arena of mRNA therapeutic development, we have upgraded the manufacturing and quality attributes of our trusted raw material reagents to ensure they meet the strict quality requirements for the manufacture of mRNA therapeutics.
Related Products
Similar Products
Ribo m7G Cap Analog
Enhances translation efficiency in many reticulocyte-based reactions.
P1711, P1712
TLR Reporter Bioassay
Measure the potency and stability of TLR agonists.
JA9011, JA9015, GA1322, GA6010
QuantiFluor® RNA System
Fluorescent quantitation of small amounts of RNA in solution.
E3310
FuGENE® SI Transfection Reagent
Innovative transfection reagent for the delivery of siRNA, miRNA and other small RNA molecules into a wide range of eukaryotic cell lines.
E9311, E9312
Frequently Used With
GloMax® Discover System
High-performance microplate reader for detecting luminescence, fluorescence and absorbance.
GM3000
RiboMAX™ Large-Scale RNA Production Systems
Compatible with modified or unmodified nucleotides. Produces 2–5mg/ml RNA.
P1280, P1300
Lumit® dsRNA Detection Assay
A novel technique to quantify dsRNA in a sample; based on NanoBiT® luciferase complementation.
W2041, W2042, W2010, W2040
Maxwell® RSC simplyRNA Cells Kit
Purifies RNA from cells using a low elution volume and the Maxwell® RSC Instrument.
AS1390
