Apc antitlr4

Manufactured by Thermo Fisher Scientific

Sourced in United States, Italy

The APC-antiTLR4 is a fluorescently labeled antibody that binds to the Toll-like receptor 4 (TLR4) on the surface of cells. This product is used for the detection and analysis of TLR4 expression in flow cytometry applications.

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2 protocols using apc antitlr4

Comparative Analysis of Synovial Fibroblasts in RA

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OASFs, tOASFs, and RASFs were cultured with DMEM containing 1% heat-inactivated FBS for 24 h, and then the media were replaced with DMEM containing 10% heat-inactivated FBS and cultured for additional 24 h. Cells were collected, fixed, and stained with PI (Keygentec, Nanjing, China), and cell cycle was analyzed using a FACSCalibur with ModFit software (Becton Dickinson, Sparks, MD, USA).
OASFs, tOASFs, and RASFs were pretreated with apoptosis-inducers A (Apopisa) and B (Apobid) (1 : 1000, Beyotime, Nantong, China) for 8 h, harvested, stained with Annexin V-FITC/PI (Bender MedSystems, Vienna, Austria), and analyzed by flow cytometry.
OASFs, tOASFs, and RASFs were stained with PE-antiTLR2 (eBioScience, San Diego, CA, USA), APC-antiTLR4 (eBioScience), and anti-RAGE (Millipore, Billerica, MA, USA). The expression of HMGB1-associcated receptors was analyzed by flow cytometry.
OASFs, tOASFs, and RASFs were stained with PerCP-antiCXCL12 (R&D), APC-antiCCL2, PE-antiVCAM-1, and APC-antiICAM-1 (eBioscience). The expression of RA-associated chemokines and intercellular adhesion molecules was analyzed by flow cytometry.

Qin Y., Chen Y., Wang W., Wang Z., Tang G., Zhang P., He Z., Liu Y., Dai S.M, & Shen Q. (2014). HMGB1–LPS complex promotes transformation of osteoarthritis synovial fibroblasts to a rheumatoid arthritis synovial fibroblast-like phenotype. Cell Death & Disease, 5(2), e1077-.

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Immunophenotyping of Immune Cell Subsets

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Cells were immunostained with the following monoclonal anti-human antibodies: FITC anti-Lin3, V450 anti-CD56, APC-H7 anti-CD45, APC anti-IFN-γ, PE anti-TNF-α, PE Annexin V, 7AAD (all from Becton Dickinson, Milan, Italy), PercpCy5.5-CD127, PE anti-TLR3, PE anti-TLR9, APC anti-TLR4, PE anti-TLR2, APC anti-ROR-γt, PE anti-ROR-γt, PECy7 anti-T-bet, PE anti-IL-17A, AlexaFluor647 anti-IL-17A (all from eBioscience), PE anti-CRTH2 (Biolegend, San Diego, CA) and PE anti-TLR7 (R&D Systems). Cells were immunostained with the following anti-mouse antibodies: FITC antiCD90.2, hematopoietic lineage cocktail efluor 450, PECy7 anti-T-bet, APC anti-ROR-γt, PE anti-TNF-αPE anti-IL-17A (all from eBioscience), APC-Cy7 anti-CD45, PE anti-ROR-γt, PECy7 anti-IFN-γ (all from Becton Dickinson). In all experiments, appropriate isotype control IgGs (Becton Dickinson and eBioscience) and fluorescence minus one controls were used. All antibodies were used at 1:100 final dilution. For intracellular immunostaining, cells were fixed and permeabilized using staining buffer set and permeabilization buffer (both from eBioscience) according to the manufacturer’s instruction. Cells were analyzed by flow cytometry (FACSverse, BD Bioscience, San Jose, CA).

Marafini I., Monteleone I., Di Fusco D., Cupi M.L., Paoluzi O.A., Colantoni A., Ortenzi A., Izzo R., Vita S., De Luca E., Sica G., Pallone F, & Monteleone G. (2015). TNF-α Producing Innate Lymphoid Cells (ILCs) Are Increased in Active Celiac Disease and Contribute to Promote Intestinal Atrophy in Mice. PLoS ONE, 10(5), e0126291.

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