Lta bs

Manufactured by InvivoGen

Sourced in United States

The LTA-BS is a laboratory equipment product. It is a device designed for scientific research purposes. The core function of the LTA-BS is to perform specific tasks related to laboratory experiments and analyses.

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B. subtilis LTA (2 citations)

4 protocols using lta bs

Luciferase Assay for Cellular Signaling

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Cells were seeded in 96-well plates a day before treatment at a density of 1x10⁵ cells per ml. Cells were treated with 30 µM PAR, 100 ng/ml LTA-BS (Invivogen), 1 ng/ml LPS-B5 ultrapure (Invivogen), or 30 µM ADPr for 24 hours. Luciferase activity was determined using the Promega Luciferase Assay system according to manufacturer’s instructions.

Krukenberg K.A., Kim S., Tan E.S., Maliga Z, & Mitchison T.J. (2015). Extracellular poly(ADP-ribose) is a pro-inflammatory signal for macrophages. Chemistry & biology, 22(4), 446-452.

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Evaluating Cell Barrier Function Modulation

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DON (Sigma, Missouri, USA) was diluted in absolute ethanol to prepare 2 μg/mL solution and treated to the apical side of cell monolayer for 48 h. To evaluate the effect of whole bacteria and TLR2 agonist on the barrier function, IPEC-J2 cells were pretreated with 1×10⁹ cfu/mL of heat-inactivated B. subtilis (inactivation confirmed after the plating, data not shown), 10 μg/mL of lipoteichoic acid from B. subtilis (LTA-BS; Invivogen, San Diego, USA) or media for 1 h before the DON treatment.

Gu M.J., Song S.K., Park S.M., Lee I.K, & Yun C.H. (2014). Bacillus subtilis Protects Porcine Intestinal Barrier from Deoxynivalenol via Improved Zonula Occludens-1 Expression. Asian-Australasian Journal of Animal Sciences, 27(4), 580-586.

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Investigating TLR2 Agonists on Intestinal Barrier

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IPEC-J2 cell monolayer was treated with 2 μg/mL of DON (Sigma, Missouri, USA) for 24, 48 or 72 h. To evaluate the effect of TLR2 agonists on the barrier function, IPEC-J2 cells were pretreated with 10 μg/mL of LTA from B. subtilis (LTA-BS; Invivogen, San Diego, USA), PGN from B. subtilis (PGN-BS; Invivogen), Pam3CSK4 (Pam3Cys-SKKKK; Invivogen) or complete medium as a control for 24 h before DON treatment. In some experiments, 10 μg/mL of the PI3K inhibitor LY294002 (Cell signaling, Massachusetts, USA) or 20 μg/mL of anti-TLR2 neutralizing antibody (eBioscience, San Diego, USA) was treated prior to the treatment with TLR2 ligands.

Gu M.J., Song S.K., Lee I.K., Ko S., Han S.E., Bae S., Ji S.Y., Park B.C., Song K.D., Lee H.K., Han S.H, & Yun C.H. (2016). Barrier protection via Toll-like receptor 2 signaling in porcine intestinal epithelial cells damaged by deoxynivalnol. Veterinary Research, 47, 25.

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Comprehensive Phenotypic Profiling of Immune Cells

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The following reagents were procured as described: CD11c-FITC, CD11c-PE, CD11b-APC, I MHC class II (I-Ad/I-Ed)-FITC, CD40-V421 (BD Bioscience, USA), CD80-FITC, CD86-PE, CD206-PE, CD14-PE, CD115-PE, F4/80-APC, CD103-PE, CD8a-V430, CD197-PE, B220-PE, CD207-PE, DEC-205-PE, CD124-PE, sigH-PE (eBioscience, USA). Blocking antibodies against TRAIL, FasL, TNFR1 (Biolegend, USA), ultrapure mouse rIFNβ, rTNFα (Biolegend, USA). DAPI, Cell Trace Violet (Invitrogen, USA), L-NMMA, β-Mercaptoethanol (Sigma, USA), FeTPPS (Calbiochem, USA), Bx795 (Invitrogen, USA). LPS 055:B5 (TLR tested, Invivogen, USA). LTA-BS, Pam2CSK4, Pam3CSK4, Poly(I:C) HMW, Flagellin-PA Ultrapure, Imiquimod, ODN CpG 1826 (Invivogen, USA). Pharmaceutical grade Immunomax® (IMM) has been purchased from Immapharma Ltd (Russia).

Bagaev A., Pichugin A., Nelson E.L., Agadjanyan M.G., Ghochikyan A, & Ataullakhanov R.I. (2018). Anti-cancer mechanisms in two murine bone marrow derived-DC subsets activated with Toll-like receptor 4 agonists. Journal of immunology (Baltimore, Md. : 1950), 200(8), 2656-2669.

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