cb2 rabbit anti-mouse primary antibody  (Cayman Chemical)

Journal: International Journal of Molecular Sciences

Article Title: Beta-Caryophyllene Modifies Intracellular Lipid Composition in a Cell Model of Hepatic Steatosis by Acting through CB2 and PPAR Receptors

doi: 10.3390/ijms24076060

Figure Lengend Snippet: Effect of CB2, PPARα and PPARγ receptors antagonists on triglyceride accumulation per cell. Cells were incubated with different concentrations of BCP and 0.5 mM FFAm, in the presence or absence of specific receptor antagonists for 24 h. ( A ) Treatment of HepG2 cells with 5 µM CB2 antagonist AM630. ( B ) Treatment with 100 nM PPARα antagonist GW6471. ( C ) Treatment with 10 µM PPARγ antagonist GW9662. Data are expressed as a percentage change with respect to 0.5 mM FFA control condition (set equal to 100) and represent the mean ± SEM of five independent experiments. ** p < 0.01; *** p < 0.001 vs. BCP + 0.5 mM FFAm treated cells.

Article Snippet: Chemicals used were: (E)-β-caryophyllene (BCP) purchased from Sigma-Aldrich (St. Louis, MO, USA), AdipoRedTM assay reagent from Lonza (Walkersville, MD, USA), NucBlue Live ReadyProbes Reagent from Invitrogen (Carlsbad, CA, USA), CellTiter-Glo ® Luminescent Cell Viability from Promega (Madison, WI, USA), anti-CB2 primary antibody from Cayman chemical (Ann Arbor, MI, USA), anti-Rabbit IgG AlexaFluor647 secondary antibody from Jackson Immunoresearch (Ely, UK), AM630, GW9662 and GW6741 antagonists from Cayman chemical and sodium oleate, sodium palmitate and bovine serum albumin from Sigma-Aldrich.

Techniques: Incubation, Control

Journal: International Journal of Molecular Sciences

Article Title: Beta-Caryophyllene Modifies Intracellular Lipid Composition in a Cell Model of Hepatic Steatosis by Acting through CB2 and PPAR Receptors

doi: 10.3390/ijms24076060

Figure Lengend Snippet: qRT-PCR analysis of CB2 ( A ), PPARα ( B ) and PPARγ ( C ) mRNAs normalized for the housekeeping gene β-actin. Data are represented as the mean ± SEM of three independent experiments. * p < 0.05, *** p < 0.001 vs. control (untreated cells, set equal to 1) or vs. 0.5 mM FFAm-treated cells.

Article Snippet: Chemicals used were: (E)-β-caryophyllene (BCP) purchased from Sigma-Aldrich (St. Louis, MO, USA), AdipoRedTM assay reagent from Lonza (Walkersville, MD, USA), NucBlue Live ReadyProbes Reagent from Invitrogen (Carlsbad, CA, USA), CellTiter-Glo ® Luminescent Cell Viability from Promega (Madison, WI, USA), anti-CB2 primary antibody from Cayman chemical (Ann Arbor, MI, USA), anti-Rabbit IgG AlexaFluor647 secondary antibody from Jackson Immunoresearch (Ely, UK), AM630, GW9662 and GW6741 antagonists from Cayman chemical and sodium oleate, sodium palmitate and bovine serum albumin from Sigma-Aldrich.

Techniques: Quantitative RT-PCR, Control

Journal: International Journal of Molecular Sciences

Article Title: Beta-Caryophyllene Modifies Intracellular Lipid Composition in a Cell Model of Hepatic Steatosis by Acting through CB2 and PPAR Receptors

doi: 10.3390/ijms24076060

Figure Lengend Snippet: Localization of CB2 receptors. Representative confocal images showing CB2 immunostaining (magenta) in HepG2 cell line. Nuclei are stained with DAPI (blue). Pictures are shown as max z-projections (low magnification; left ) with white arrows and arrowheads to highlight CB2high+ cells and CB2low+ cells, respectively, and a single confocal plane with reslicing ( right ) to better appreciate the intracellular distribution of CB2+ puncta in two of the cells present in the image. The cyan contoured image shows a cell positive for CB2 at low levels; the yellow contoured image identifies a cell with extensive immunolabelling for CB2. Scale bars: 50 µm (low magnification) and 10 µm (high magnification).

Article Snippet: Chemicals used were: (E)-β-caryophyllene (BCP) purchased from Sigma-Aldrich (St. Louis, MO, USA), AdipoRedTM assay reagent from Lonza (Walkersville, MD, USA), NucBlue Live ReadyProbes Reagent from Invitrogen (Carlsbad, CA, USA), CellTiter-Glo ® Luminescent Cell Viability from Promega (Madison, WI, USA), anti-CB2 primary antibody from Cayman chemical (Ann Arbor, MI, USA), anti-Rabbit IgG AlexaFluor647 secondary antibody from Jackson Immunoresearch (Ely, UK), AM630, GW9662 and GW6741 antagonists from Cayman chemical and sodium oleate, sodium palmitate and bovine serum albumin from Sigma-Aldrich.

Techniques: Immunostaining, Staining

Journal: Pharmaceutics

Article Title: Cannabinoid Receptor 2 (CB2) Inverse Agonist SMM-189 Induces Expression of Endogenous CB2 and Protein Kinase A That Differentially Modulates the Immune Response and Suppresses Experimental Colitis

doi: 10.3390/pharmaceutics14050936

Figure Lengend Snippet: Induction of CB2, PKA, and histological alteration after SMM-189 treatment. LPLs and colon tissue were harvested from the three groups of mice in this study. ( A ) represents the immunoblot analysis of CB2 and PKA. ( B ) represents the relative fold expression of CB2 and ( C ) represents the relative fold expression of PKA. ( D ) representative histological sections of colons from the three groups of mice. DSS-treated mice that did not receive SMM-189 treatment exhibited significant lymphocyte infiltration and distortion of crypts ( central panel arrow), but SMM-189-treated mice exhibited markedly decreased lymphocyte infiltration ( right panel); these changes were not observed in tissue from mice in the control group ( left panel). Other changes include diffuse leukocyte infiltrates and thickening of the lamina propria near the distorted crypts in the colon. ( E ) represents the combined inflammation scores, which were reduced for the SMM-189-treated group compared to DSS alone. Representative sections from three separate experiments (10X magnification) are shown, with each group containing six mice ( n = 18). Asterisks (*) indicate statistically significant differences at p < 0.01 between the vehicle (DSS) and SMM-189-treated groups.

Article Snippet: The membrane was blocked with intercept blocking buffer (#92760001, LI-COR Biosciences, Lincoln, NE, USA) at room temperature (RT) for 1 h and incubated with mouse primary antibodies anti-CB2 (#SC-293188, Santa Cruz Biotechnology, Dallas, TX, USA; 1:200), anti-PKA (#SC-136231, Santa Cruz Biotechnology; 1:200), anti-β-actin (#926-42212, LI-COR Biosciences; 1:1000) at 4 °C overnight while shaking.

Techniques: Western Blot, Expressing, Control