TLR4/HEK293 Stable Cell Line

TLR4/HEK293 Stable Cell Line is a stably transfected HEK293 cell line which expresses human Toll-like receptor 4 (TLR4, also designated as CD284) . TLR4 mediates immune responses by recognition of lipoplysaccharides (LPS) with the help of LPS-binding protein, CD14 and MD-2. TLR4 stimulation uniquely triggers two signaling pathways named the MyD88-dependent pathway and the TRIF-dependent pathway, which lead to the production of two sets of cytokines such as proinflammatory cytokines and type I interferons/chemokines, respectively. Note that TLR4 in the TLR4/HEK293 stable cell line contains the N-terminal HA tag (Figure 2), which does not interfere with TLR4 activity as confirmed by functional assay (Figure 1) . Sequence data: Human TLR4 (accession number NP_612564) MMSASRLAGTLIPAMAFLSCVRPESWEPCVEVVPNITYQCMELN FYKIPDNLPFSTKNLDLSFNPLRHLGSYSFFSFPELQVLDLSRCEIQTIEDGAYQSLS HLSTLILTGNPIQSLALGAFSGLSSLQKLVAVETNLASLENFPIGHLKTLKELNVAHN LIQSFKLPEYFSNLTNLEHLDLSSNKIQSIYCTDLRVLHQMPLLNLSLDLSLNPMNFI QPGAFKEIRLHKLTLRNNFDSLNVMKTCIQGLAGLEVHRLVLGEFRNEGNLEKFDKSA LEGLCNLTIEEFRLAYLDYYLDDIIDLFNCLTNVSSFSLVSVTIERVKDFSYNFGWQH LELVNCKFGQFPTLKLKSLKRLTFTSNKGGNAFSEVDLPSLEFLDLSRNGLSFKGCCS QSDFGTTSLKYLDLSFNGVITMSSNFLGLEQLEHLDFQHSNLKQMSEFSVFLSLRNLI YLDISHTHTRVAFNGIFNGLSSLEVLKMAGNSFQENFLPDIFTELRNLTFLDLSQCQL EQLSPTAFNSLSSLQVLNMSHNNFFSLDTFPYKCLNSLQVLDYSLNHIMTSKKQELQH FPSSLAFLNLTQNDFACTCEHQSFLQWIKDQRQLLVEVERMECATPSDKQGMPVLSLN ITCQMNKTIIGVSVLSVLVVSVVAVLVYKFYFHLMLLAGCIKYGRGENIYDAFVIYSS QDEDWVRNELVKNLEEGVPPFQLCLHYRDFIPGVAIAANIIHEGFHKSRKVIVVVSQH FIQSRWCIFEYEIAQTWQFLSSRAGIIFIVLQKVEKTLLRQQVELYRLLSRNTYLEWE DSVLGRHIFWRRLRKALLDGKSWNPEGTVGTGCNWQEATSI

Product Specifications

Applications

Functional Assay

Components

Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO

Shipping Conditions

Dry Ice

Storage Conditions

Immediately upon receipt, store in liquid nitrogen.

Applications Notes

Application:. Functional assay. Culture conditions: Cells should be grown at 37oC with 5% CO2 using DMEM medium (w/ L-Glutamine, 4.5g/L Glucose and Sodium Pyruvate) supplemented with 10% heat-inactivated FBS and 1% Pen/Strep, plus 10 μg/ml of Blasticidin. It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37oC water-bath, transfer to a tube containing 10 ml of growth medium without Blasticidin, spin down cells, resuspend cells in pre-warmed growth medium without Blasticidin, transfer resuspended cells to T25 flask and culture in 37oC-CO2 incubator. Leave the T25 flask in the incubator for 1~2 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 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. To achieve satisfactory results, cells should not be passaged over 16 times. LIMITED USE RESTRICTIONS: THIS PRODUCT IS SOLELY FOR IN VITRO RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE. By use of this product, user agrees to be bound by the terms of this limited use statement. This product is solely for Internal Research Purposes and not for Commercial Purposes. Commercial Purposes include, but are not limited to (1) use of the cell line in manufacturing; (2) use of the cell line to provide a service, information or data; (3) use of the cell line for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the cell line whether or not such cell lines are resold for use in research. The buyer cannot sell, give or otherwise transfer this product to a third party. Commercial License Agreement is available for non-research use if applicable. Please contact Abeomics (info@abeomics.com) .