Cell lines B16F10 (ATCC® CRL-6475™), CT26 (ATCC® CRL-2638™), HEK-blue-TLR2 (Invivogen), HEK-blue-TLR3 (Invivogen), HEK-blue-TLR4 (Invivogen), HEK-blue-TLR7 (Invivogen), HEK-blue-TLR9 (Invivogen), Raw-Lucia ISG (Invivogen), A549-Dual™ Cells (Invivogen), A549-Dual™ KO-MDA5 Cells (Invivogen), A549 (ATCC® CCL-185™), Hela (ATCC® CCL-2™), and SK-MEL-5 (ATCC® HTB-70™) were cultured following vendor instructions. YUMMER 1.7 cells were a gift from Prof. Marcus Bosenberg at Yale University and followed the procedures for culture described previously44 (link). Female C57BL/6J (JAX Stock No. 000664), Balb/C (JAX Stock No. 000651), Pmel Thy1.1 (JAX Stock No. 005023), MyD88−/− (JAX Stock No. 009088), STING−/− (Jax Stock No. 025805), and Batf3−/− (JAX Stock No. 013755) mice 6–8 weeks of age were purchased and maintained in the animal facility at the Massachusetts Institute of Technology (MIT). All animal studies and procedures were carried out following federal, state, and local guidelines under an IACUC-approved animal protocol by Committee of Animal Care at MIT.
Hek blue tlr4
Manufactured by InvivoGen
Sourced in United States
HEK-Blue™ TLR4 is a reporter cell line that expresses the human Toll-like receptor 4 (TLR4) and an NF-κB-inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene. The activation of TLR4 by its ligands induces the activation of NF-κB, leading to the secretion of SEAP, which can be easily detected in the cell culture supernatant.
Automatically generated - may contain errors
Lab products found in correlation
Hek blue null1 v, by InvivoGen (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Hek blue sr ai, by Sino Biological (2 mentions)
Hek blue cd36, by Sino Biological (2 mentions)
Hek blue null1 v, by Sino Biological (2 mentions)
Pcmv hygro sr ai, by Sino Biological (2 mentions)
Pcmv hygro cd36, by Sino Biological (2 mentions)
Thp 1 monocytic mφ cell line, by American Type Culture Collection (2 mentions)
Sk mel 5, by American Type Culture Collection (1 mentions)
Hek blue tlr2, by InvivoGen (1 mentions)
Hek blue tlr3, by InvivoGen (1 mentions)
Hek blue tlr7, by InvivoGen (1 mentions)
Hek blue tlr9, by InvivoGen (1 mentions)
Raw lucia isg, by InvivoGen (1 mentions)
A549 dual cells, by InvivoGen (1 mentions)
Mle 12 cells, by American Type Culture Collection (1 mentions)
Thp 1 cell line, by American Type Culture Collection (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
Dnase 1, by Qiagen (1 mentions)
Nigericin, by Merck Group (1 mentions)
5 protocols using hek blue tlr4
1
Cell Line and Mouse Model Protocols
2
Culturing and Differentiation of Murine Macrophages
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HEK-Blue™ TLR4 (InvivoGen, San Diego, CA, USA) and L929 (ATCC, Manassas, Virginia, USA) cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS, HyClone, Logan, Utah, USA) and 1× antibiotics (Gibco™, Massachusetts, USA). MLE-12 cells (ATCC) were maintained in DMEM supplemented with 2% FBS and 1× antibiotics. No mycoplasma contamination was detected. To prepare BMDMs, bone marrow cells were harvested from the femur and tibia of C57BL/6 N and TLR4 knockout mice and differentiated in DMEM containing 25% L929-conditioned medium, 10% FBS, and 1× antibiotics. L929-conditioned medium was obtained by culturing L929 cells in DMEM for 7 days until a confluence of >90% was obtained. Afterwards, the medium underwent centrifugation at 1500 rpm for 10 min and the supernatants were collected and filtered through a 0.45 μm filter system.
3
Macrophage THP-1 cell line protocol
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The human macrophage THP-1 cell line was from the American Type Culture Collection (Cat# TIB-202, RRID: CVCL_0006). HEK-Blue™ TLR4 (Cat# hkb-mtlr4; RRID: CVCL_IM89) and RAW-Blue™ cells (Cat# Raw-sp; RRID: CVCL_X594), Pam3csK4, and anti-TLR2 (Cat# mab2-mtlr2; RRID: AB_1277471 ) were from InvivoGen (San Diego, USA). E. coli lipopolysaccharide (LPS) 055:B5, phorbol 12-myristate 13-acetate (PMA), nigericin, MCC950, cytochalasin D, monodansylcadaverine, N-acetyl-L-cysteine, ATP and A438079 were from Sigma–Aldrich (San Luis, USA). Ac-YVAD-AOM and mitoTEMPO were from Merck-Millipore (Burlington, USA). Proteinase K was from Roche (Basel, Switzerland), DNase I from Qiagen (Hilden, Germany), Uricase from Worthington (Lakewood, USA), anti-HMGB1 (Cat# 651,401; RRID: AB_10945159 ) from Biolegend (San Diego, USA), and recombinant human IL-18 binding protein (IL-18BP) from GeneScript (Piscataway, USA).
4
THP-1 and HEK-Blue Cell Culture Protocols
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Cell Culture Experiments a) The THP-1 monocytic MΦ cell line was obtained from American Type Culture Collection (ATCC# TIB-202; Manassas, VA) and incubated at 37°C and 5% CO 2 in complete RPMI 1640 growth medium supplemented with 10 U/ml penicillin and streptomycin, 1mM sodium pyruvate (11965092; Thermo Fisher Scientific, Burlington, ON) and heat inactivated 10% Fetal Bovine Serum (FBS; Gibco, Burlington, ON). These undifferentiated mononuclear cells (between passage 3 and 16) were seeded in 24-well plates at concentration of 1×10 6 cells / well, and stimulated with 50 ng/ml phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, Oakville, ON) to induce differentiation to MΦ-like cells (12) . b) HEK-Blue TM Null1-v and HEK-Blue TM -TLR4 (stably expressing TLR4, MD-2 and CD14 coreceptor genes) cell lines were obtained from Invivogen (SanDiego, CA, USA). HEK-Blue TM SR-AI and HEK-Blue TM CD36 were prepared by stably transfecting the parental cell line HEK-Blue TM Null1-v with plasmid pCMV/Hygro-SR-AI or pCMV/Hygro-CD36 (Sino Biological Inc., North Wales, PA) and selecting using the addition of 100 μg/mL hygromycin.
5
THP-1 and HEK-Blue Cell Culture Protocols
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Cell Culture Experiments a) The THP-1 monocytic MΦ cell line was obtained from American Type Culture Collection (ATCC# TIB-202; Manassas, VA) and incubated at 37°C and 5% CO 2 in complete RPMI 1640 growth medium supplemented with 10 U/ml penicillin and streptomycin, 1mM sodium pyruvate (11965092; Thermo Fisher Scientific, Burlington, ON) and heat inactivated 10% Fetal Bovine Serum (FBS; Gibco, Burlington, ON). These undifferentiated mononuclear cells (between passage 3 and 16) were seeded in 24-well plates at concentration of 1×10 6 cells / well, and stimulated with 50 ng/ml phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, Oakville, ON) to induce differentiation to MΦ-like cells (12) . b) HEK-Blue TM Null1-v and HEK-Blue TM -TLR4 (stably expressing TLR4, MD-2 and CD14 coreceptor genes) cell lines were obtained from Invivogen (SanDiego, CA, USA). HEK-Blue TM SR-AI and HEK-Blue TM CD36 were prepared by stably transfecting the parental cell line HEK-Blue TM Null1-v with plasmid pCMV/Hygro-SR-AI or pCMV/Hygro-CD36 (Sino Biological Inc., North Wales, PA) and selecting using the addition of 100 μg/mL hygromycin.
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