PCL (Mw=80 kDa), pluronic-F-127, gelatin, sodium borohydride, and Triton X-100 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dichloromethane (DCM) and N, N-dimethylformamide (DMF) were purchased from BDH Chemicals (Dawsonville, GA, USA). Dulbecco’s modified eagle medium (DMEM), fetal bovine serum (FBS), and penicillin-streptomycin were obtained from Invitrogen (Carlsbad, CA, USA). CD 31, Ki67, TNF-α, IL-4, IL-6, IL-10 primary antibodies and Goat Anti-Rabbit IgG H&L (FITC), Goat Anti-Mouse IgG H&L (Alexa Fluor® 647), Goat Anti-Rat IgG H&L (Cy5®), Donkey Anti-Goat IgG H&L (Cy5®) secondary antibodies, phalloidin-iFluor 488 reagent (ab176753) were purchased from Abcam (Cambridge, MA, USA). CD206 and CCR7 primary antibodies were purchased from NovusBio (Centennial, CO, USA). The Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies, Inc (Rockville, MD, USA).
Cd206
Manufactured by Novus Biologicals
Sourced in United States
CD206, also known as the macrophage mannose receptor, is a transmembrane glycoprotein expressed on the surface of macrophages and other antigen-presenting cells. It functions as an endocytic receptor, mediating the internalization of glycosylated ligands for degradation or presentation.
Automatically generated - may contain errors
Lab products found in correlation
Dichloromethane dcm, by Avantor (1 mentions)
Pcl mw 80 kda, by Merck Group (1 mentions)
Phalloidin ifluor 488 reagent, by Abcam (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Sodium borohydride, by Merck Group (1 mentions)
Gelatin, by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Pluronic f 127, by Merck Group (1 mentions)
Goat anti rabbit igg h l, by Abcam (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Alexa 555 secondary antibodies, by Thermo Fisher Scientific (1 mentions)
Lsm 900, by Zeiss (1 mentions)
Cd11b, by Merck Group (1 mentions)
Anti nf κb p65 mouse monoclonal antibody, by Santa Cruz Biotechnology (1 mentions)
Osmium tetroxide, by Electron Microscopy Sciences (1 mentions)
Cd11b, by Novus Biologicals (1 mentions)
Col1a1, by Abcam (1 mentions)
S 4700, by Hitachi (1 mentions)
Saponin, by Thermo Fisher Scientific (1 mentions)
Dp80 microscope, by Olympus (1 mentions)
5 protocols using cd206
1
Bioactive Hydrogel Scaffold Synthesis
2
Comprehensive Wound Healing Analysis
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The wounds and a 5 mm unwounded skin border were harvested from mice and processed for histological analysis. Wound widths were measured as the distance between wound margins. Wound areas were determined using image analysis (measured below the clot and above the panniculus muscle). Re-epithelialization was defined as the percentage of distance migrated by the neoepidermis compared to the upper wound width. Immunohistochemistry analysis was performed on formalin-fixed, paraffin-embedded 5-μm sections. The primary antibodies used in this study were against keratin 14 (K14, 1:1000, Covance, Princeton, NJ, USA), Ki67 (1:1000, Covance) and CD206 (1:500, Novus Biologicals, CO, USA). To quantify CD206 expression, ImageJ 1.50d (USA) was used to analyze the integrated optical density.
3
Microglial Cell Immunocytochemistry on Chip
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Microglial cells on the microfluidic chip were fixed with 4% paraformaldehyde and permeabilized with 0.2% Triton X-100 in phosphate-buffered saline (PBS; Gibco-BRL, Gaithersburg, MD, USA) for 15 min at room temperature (RT). Blocking was performed using 3% bovine serum albumin (BSA; Millipore, MN, USA) in PBS (1% w/v) for approximately 2 h at RT. Subsequently, the cells were incubated with CD11b (Millipore, MN, USA) and CD206 (Novus biologicals, CO, USA) primary antibodies for 2 h at 37 °C at a dilution ratio of 1:100 and washed with PBS containing 1% BSA. Thereafter, the cells were incubated with Alexa 555 secondary antibody (1:200; Invitorgen, Waltham, MA, USA) for 2 h at 37 °C.
Human NP cells were treated with recombinant IL-1β for 48 h and fixed and blocked with proprietary reagents. Anti-NF-κB p65 mouse monoclonal antibody (Santa Cruz, Dallas, TX, USA) was used to detect NF-κB p65 protein. Goat anti-mouse Alexa 555 secondary antibodies (Invitrogen, Waltham, MA, USA) and 5% BSA were used for secondary incubation in PBS for 1 h at room temperature. After staining with 4′,6-diamidino-2-phenylindole (DAPI) for 10 min, fluorescence images were acquired using a confocal microscope (Zeiss, LSM 900).
Human NP cells were treated with recombinant IL-1β for 48 h and fixed and blocked with proprietary reagents. Anti-NF-κB p65 mouse monoclonal antibody (Santa Cruz, Dallas, TX, USA) was used to detect NF-κB p65 protein. Goat anti-mouse Alexa 555 secondary antibodies (Invitrogen, Waltham, MA, USA) and 5% BSA were used for secondary incubation in PBS for 1 h at room temperature. After staining with 4′,6-diamidino-2-phenylindole (DAPI) for 10 min, fluorescence images were acquired using a confocal microscope (Zeiss, LSM 900).
4
Comprehensive Tissue Characterization via Histology and SEM
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Formalin-fixed paraffin-embedded tissues samples were sectioned with 5-μm thicknesses and stained with H&E, SR, and immunostaining for alpha-smooth muscle actin (α-SMA, ACTA2) (1:100, Sigma-Aldrich St. Louis, MO) and collagen, Type 1, alpha 1 (COL1A1) (1:200, Abcam, Cambridge, UK), Pxdn (provided by Dr Gutam, 1:200), and CD11b (1:100, Novus Biologicals, Centennial, CO), CD206 (1:100, Novus Biologicals) according to standard protocols, as reported previously.70 (link) For tissue analysis using the SEM, formalin-fixed and paraffin-embedded sections were incubated in osmium tetroxide (0.5%; Electron Microscopy Sciences, 19152) before serial dehydration in ethanol. Samples were then dried using a critical point drier and imaged using field emission SEM (Hitachi S-4700).
5
Immunofluorescence Analysis of Polarized TAMs
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THP–1 cells were plated on glass coverslips, polarized to TAMs, and given drug treatments as described above. Upon treatment, the coverslips were washed with 1X PBS, and cells were fixed in 4% PFA for 20 minutes at RT. The cells were permeabilized with 0.1% Triton X-100 for 15 minutes at RT and blocked with IF buffer (1.3M NaCl, 70mM Na2HPO4, 35mM NaH2PO4, 77mM NaN3, 1% BSA, 2% Triton X-100 and 5% Tween-20) containing 10% goat serum for 1 hour. Cells were washed with PBS and incubated overnight at 4°C with primary antibody solution: Antibody in IF buffer containing 3% Saponin (Fisher Scientific, Cat. No. 55–825-5100GM). Antibodies used were as follows: TNFα (ThermoFisher, Cat. No. MA523720), NOS2 (ThermoFisher, Cat. No. PA1–036), CD163 (Abcam, Waltham, MA, Cat. No. ab182422), CD206 (Novus Biologicals, Centennial, CO, Cat. No. NPB1–90020), IL10 (R&D systems, Minneapolis, MN, Cat. No. MAB9184–100) and Dectin 1 (ThermoFisher, Cat. No. PA583996). Cells were then washed with PBS twice and incubated with secondary antibody solution (1:200 dil) for 2 hours followed by DAPI stain for 15 minutes at RT. Coverslips were then mounted onto slides as described above. Fluorescence images were captured using Olympus DP80 microscope and analyzed using ImageJ software.
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