Paeoniflorin (PF, >95 % purity), DAPI fluorescent stain, and conA type IV were obtained from Sigma (St Louis, MO, USA). The SABC kit for immunohistochemical analysis was obtained from Boster (Wuhan, China). The IL1β ELISA kit was from R&D system (Minneapolis, MN, USA). The antibodies used for the immunohistochemical and western blot analyses were rabbit polyclonal IL1β (sc-7884), goat polyclonal monocyte chemotactic protein 1 (MCP1) (sc-1785), rabbit polyclonal F4/80 (sc-25830), mouse monoclonal CXCR3 (sc-137140) and rabbit polyclonal CXCL11 (sc-28874) purchased from Santa Cruz Biotechnology (La Jolla, CA, USA). Mouse monoclonal CD68 (MCA31R) was obtained from Serotec (Oxfordshire, OX51GE, UK). Secondary fluorescence-labelling goat anti-mouse Cy3 and goat anti-rabbit FITC second antibodies were obtained from Jackson (West Grove, PA, USA).
Cona type 4
Manufactured by Merck Group
Sourced in United States
ConA (type IV) is a laboratory equipment product manufactured by Merck Group. It is a type of lectin, a protein that binds to specific carbohydrates. ConA (type IV) is commonly used in various biological and biochemical applications.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse cy3, by Jackson ImmunoResearch (1 mentions)
Il 1β elisa kit, by R&D Systems (1 mentions)
Paeoniflorin pf, by Merck Group (1 mentions)
Dapi fluorescent stain, by Merck Group (1 mentions)
Suramin, by Merck Group (1 mentions)
Concanavalin a (cona), by Merck Group (1 mentions)
Alexa 647, by Thermo Fisher Scientific (1 mentions)
Alexa 350, by Thermo Fisher Scientific (1 mentions)
Alexa 488, by Thermo Fisher Scientific (1 mentions)
Sulfo cy7 nhs ester, by Lumiprobe (1 mentions)
Tcrαβ, by Thermo Fisher Scientific (1 mentions)
Ox 40l, by R&D Systems (1 mentions)
Mouse t cell enrichment column, by R&D Systems (1 mentions)
Cd40l, by R&D Systems (1 mentions)
Dri chem 3500i analyzer, by Fujifilm (1 mentions)
6 protocols using cona type 4
1
Paeoniflorin Modulates Inflammatory Pathways
2
Concanavalin A-Induced Liver Injury Model
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ConA (type IV) was purchased from Sigma‐Aldrich (St Louis, MO). Phosphate‐buffered saline or ConA (20 mg/kg) was intravenously administered to the tail vein of euthanized mice 12 hours before tissue harvest. In some experiments, RAG2 KO mice received ConA followed by intraperitoneal treatment with recombinant IFN‐γ (rIFN‐γ; 500 ng) and recombinant TNF (rTNF; 500 ng) as reported.(25 (link)
) After being fed a Cont‐ or Inu‐diet for 2 weeks, some mice were coinjected with ConA and the purinergic P2 receptor antagonist suramin (Sigma‐Aldrich; 200 μg/g body weight dissolved in pure water, intraperitoneally). Some Cont‐ or Inu‐diet‐fed mice were intravenously administered plasma of portal vein blood (200 μL/mouse) derived from ConA‐treated mice 2 hours before ConA administration, as described.(26
)
) After being fed a Cont‐ or Inu‐diet for 2 weeks, some mice were coinjected with ConA and the purinergic P2 receptor antagonist suramin (Sigma‐Aldrich; 200 μg/g body weight dissolved in pure water, intraperitoneally). Some Cont‐ or Inu‐diet‐fed mice were intravenously administered plasma of portal vein blood (200 μL/mouse) derived from ConA‐treated mice 2 hours before ConA administration, as described.(26
)
3
ConA-Induced Mouse Liver Injury
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ConA type IV (Sigma-Aldrich) was injected i.v. into the tail vein of mice (6- to 8-week-old males) 18 hours before the study endpoint at a dose of 15 mg/kg.
4
Conjugation of Concanavalin A with Cy7
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For cell wall staining, we purchased Con A conjugated with Alexa 350, Alexa 488, TRITC, and Alexa 647 from Life Technologies. Stock solutions with concentration 2.5 mg/ml were prepared in PBS and stored at −20°C in small aliquots. Con A conjugated to Cy7 stock solution was prepared using the following protocol (Mund et al., 2014 (link)). Sulfo-Cy7 NHS ester (Lumiprobe) was diluted in DMSO to a concentration of 10 mM. Con A (type IV; Sigma-Aldrich) 2.5 mg/ml solution was prepared in 0.2 M NaHCO3 with pH 8.2. Dye and protein solutions were mixed 6:100 and incubated for 4 h at room temperature. Conjugated Con A was separated from the reaction on a disposable Sephadex G-25 column, and buffer was exchanged to PBS. The stock solution was stored frozen in small aliquots.
5
T-cell Activation and Cytokine Profiling
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ConA (type IV) was purchased from Sigma (St. Louis, MO, USA). Mouse T-cell enrichment columns and neutralizing antibodies against B7.1, B7.2, OX40L and CD40L were obtained from R&D Systems (Minneapolis, MN, USA). Purified antibodies against mouse CD3, CD28 and cytometric Bead Array for mouse IFN-γ were purchased from BD Biosciences. Fluorochrome-conjugated antibodies against mouse CD3, CD4, CD8, CD19, CD44, CD45.1, CD45.2, B220, NK1.1, TCRαβ and CXCR3 were obtained from eBioscience (San Diego, CA, USA). ALT Detection Kits were purchased from NanJing JianCheng Biochemical Institute (Nan Jing, Jiang Su, China).
6
Concanavalin A Liver Injury Model
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Con A (type IV) was purchased from Sigma-Aldrich (St Louis, MO). Phosphate-buffered saline (PBS) or Con A solution (20 mg/kg) was administered into the tail vein at 1, 3, 6, or 12 hours before experiments. Under anesthesia, all mice were euthanized and their serum alanine aminotransferase (ALT) levels were measured using a DRI-CHEM 3500i Analyzer (FujiFilm, Tokyo, Japan).
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