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Testing Data (Fig-1: Induction of TLR3 activity by poly(I: C) in TLR3/ISRELeeporter HEK293 cells.)

TLR3/ISRE cell line

TLR3/ISRE Leeporter HEK293 Cell Line

Gene Names
TICAM1; TRIF; IIAE6; MyD88-3; PRVTIRB; TICAM-1
Applications
Functional Assay
Synonyms
TLR3/ISRE; TLR3/ISRE Leeporter HEK293 Cell Line; TLR3/ISRE cell line
Ordering
For Research Use Only!
Form/Format
Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
Sequence Length
712
Applicable Applications for TLR3/ISRE cell line
Functional Assay
Application Notes
Monitor the TLR3 signaling pathway activity. Screen for activators or inhibitors of the TLR3 signaling pathway. Culture conditions: Cells should be grown at 37 degree C with 5% CO2 using DMEM medium supplemented with 10% FBS and 1% Pen/Strep, plus 2 ug/ml Puromycin and 5 ug/ml blasticidin. It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37 degree C water-bath, transfer to a tube containing 10 ml of growth medium without Puromycin and Blasticidin, spin down cells, resuspend cells in pre-warmed growth medium without Puromycin and Blasticidin, transfer resuspended cells to T25 flask and culture in 37 degree C-CO2 incubator. Leave the T25 flask in the incubator for 2~4 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Puromycin and Blasticidin. Cells should be split before they reach complete confluence. To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1: 10 to 1: 20 weekly. Functional validation: A. Response of TLR3/ISRE Leeporter- HEK293 cells to Poly(I: C). 1. Harvest TLR3/ISRE Leeporter- HEK293 cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 5 x 10^4 cells/well. 2. Incubate cells at 37 degree C in a CO2 incubator for overnight. 3. The next day, stimulate cells with various concentrations of poly(I: C). 4. Incubate at 37 degree C in a CO2 incubator for 6-16 hours. 5. Add 30-50 ul of luciferase assay reagent per well. 6. Incubate at room temperature for 1-5 minutes and measure luminescence using a microplate luminometer.
Shipping Note
Product available only in the USA.
Dry Ice Shipment
Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
Preparation and Storage
Immediately upon receipt, store in liquid nitrogen.

Testing Data

(Fig-1: Induction of TLR3 activity by poly(I: C) in TLR3/ISRELeeporter HEK293 cells.)

Testing Data (Fig-1: Induction of TLR3 activity by poly(I: C) in TLR3/ISRELeeporter HEK293 cells.)
Related Product Information for TLR3/ISRE cell line
The TLR3/IFNB Leeporter cell line is a stably transfected HEK 293 cell line which expresses full-length human Toll-like receptor 3 (TLR3) and Renilla luciferase reporter gene under the transcriptional control of the interferon stimulated response element (ISRE). Functional activity of the cell line has been validated by TLR3 ligand assay (Figure 1), in which upon activation by poly (I: C), TLR3 quickly initiates TRIF-dependent signaling pathway and mediates nuclear translocation of IRF3 that induces ISRE activity.
Product Categories/Family for TLR3/ISRE cell line

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
Molecular Weight
76,422 Da
NCBI Official Full Name
TIR domain-containing adapter molecule 1
NCBI Official Synonym Full Names
toll like receptor adaptor molecule 1
NCBI Official Symbol
TICAM1
NCBI Official Synonym Symbols
TRIF; IIAE6; MyD88-3; PRVTIRB; TICAM-1
NCBI Protein Information
TIR domain-containing adapter molecule 1
UniProt Protein Name
TIR domain-containing adapter molecule 1
UniProt Gene Name
TICAM1
UniProt Synonym Gene Names
PRVTIRB; TRIF; TICAM-1; MyD88-3; TIR domain-containing adapter protein inducing IFN-beta

NCBI Description

This gene encodes an adaptor protein containing a Toll/interleukin-1 receptor (TIR) homology domain, which is an intracellular signaling domain that mediates protein-protein interactions between the Toll-like receptors (TLRs) and signal-transduction components. This protein is involved in native immunity against invading pathogens. It specifically interacts with toll-like receptor 3, but not with other TLRs, and this association mediates dsRNA induction of interferon-beta through activation of nuclear factor kappa-B, during an antiviral immune response. [provided by RefSeq, Jan 2012]

Uniprot Description

TICAM1: an adaptor protein involved in innate immunity against invading pathogens. Associates with TLR3 and TLR4 (through TICAM2) to mediate NF-kappa-B and interferon-regulatory factor (IRF) activation, and to induce apoptosis. Ligand binding to these receptors results in TICAM1 recruitment through its TIR domain. Distinct protein-interaction motifs allow recruitment of the effector proteins TBK1, TRAF6 and RIPK1, which in turn, lead to the activation of transcription factors IRF3 and IRF7, NF-kappa-B and FADD respectively. Interacts with the TIR domain of TLR3. Interacts with AZI2, TBK1, IRF3 and IRF7. Interacts with TICAM2 in TLR4 recruitment. Interaction with PIAS4 inhibits the TICAM1-induced NF-kappa-B, IRF and IFNB1 activation. Interacts with IKBKB and IKBKE. Interaction with SARM1 blocks TICAM1-dependent transcription factor activation. Interacts with TRAF3. Interacts with TRAFD1.

Protein type: Adaptor/scaffold; Apoptosis

Chromosomal Location of Human Ortholog: 19p13.3

Cellular Component: cytosol; endosome membrane

Molecular Function: protein binding; protein kinase binding; signal transducer activity

Biological Process: I-kappaB kinase/NF-kappaB cascade; MyD88-independent toll-like receptor signaling pathway; negative regulation of MyD88-independent toll-like receptor signaling pathway; positive regulation of autophagy; positive regulation of I-kappaB kinase/NF-kappaB cascade; response to exogenous dsRNA

Disease: Herpes Simplex Encephalitis, Susceptibility To, 4

Research Articles on TLR3/ISRE

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Product Notes

The TLR3/ISRE ticam1 (Catalog #AAA668885) is a Cell Line and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's TLR3/ISRE can be used in a range of immunoassay formats including, but not limited to, Functional Assay. Monitor the TLR3 signaling pathway activity. Screen for activators or inhibitors of the TLR3 signaling pathway. Culture conditions: Cells should be grown at 37 degree C with 5% CO2 using DMEM medium supplemented with 10% FBS and 1% Pen/Strep, plus 2 ug/ml Puromycin and 5 ug/ml blasticidin. It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37 degree C water-bath, transfer to a tube containing 10 ml of growth medium without Puromycin and Blasticidin, spin down cells, resuspend cells in pre-warmed growth medium without Puromycin and Blasticidin, transfer resuspended cells to T25 flask and culture in 37 degree C-CO2 incubator. Leave the T25 flask in the incubator for 2~4 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Puromycin and Blasticidin. Cells should be split before they reach complete confluence. To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1: 10 to 1: 20 weekly. Functional validation: A. Response of TLR3/ISRE Leeporter- HEK293 cells to Poly(I: C). 1. Harvest TLR3/ISRE Leeporter- HEK293 cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 5 x 10^4 cells/well. 2. Incubate cells at 37 degree C in a CO2 incubator for overnight. 3. The next day, stimulate cells with various concentrations of poly(I: C). 4. Incubate at 37 degree C in a CO2 incubator for 6-16 hours. 5. Add 30-50 ul of luciferase assay reagent per well. 6. Incubate at room temperature for 1-5 minutes and measure luminescence using a microplate luminometer. Researchers should empirically determine the suitability of the TLR3/ISRE ticam1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "TLR3/ISRE, Cell Line" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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