labeling immunofluorescence analysis  (Santa Cruz Biotechnology)

Journal: Cerebral Cortex (New York, NY)

Article Title: The Hyperpolarization-Activated HCN4 Channel is Important for Proper Maintenance of Oscillatory Activity in the Thalamocortical System

doi: 10.1093/cercor/bhz047

Figure Lengend Snippet: Characterization of brain-specific HCN4-deficient mice. ( A ) HCN4 protein expression in thalamus (Th) and frontal cortex (Cx) specimens from control (Ctr) and knockout (KO) mice. MAP-Kinase (MAPK), loading control. ( B ) Analysis of the mRNA expression of the 4 HCN isoforms in the CM, the VB and the somatosensory cortex (SSC), n = 3/3 KO and control mice. Isoform expression levels were not different between control and knockout except for HCN4 ( P < 0.001). ( C ) Top, sagittal sections of control and knockout mice labeled with an HCN4-antibody (blue staining). Sections were counterstained with Nuclear Fast Red. Bottom, detection of HCN4 in frontal sections (DAB staining). ( D ) HCN4-immunofluorescence (red) in the SSC, NRT and VB. Left panels: control, right panel: HCN4-KO. Neurons were colabeled by an anti-NeuN antibody (green). Scale bar, 50 μm.

Article Snippet: Antibodies used for double-label immunofluorescence analysis were polyclonal rabbit anti-HCN4 (1:200, Alomone Labs) and monoclonal mouse anti-NeuN conjugated with Alexa Fluor-488 (1:100, Santa Cruz Biotechnology).

Techniques: Expressing, Knock-Out, Labeling, Staining, Immunofluorescence

Journal: BMB Reports

Article Title: Effects of α-lipoic acid on LPS-induced neuroinflammation and NLRP3 inflammasome activation through the regulation of BV-2 microglial cells activation

doi: 10.5483/BMBRep.2019.52.10.026

Figure Lengend Snippet: α-LA inhibits the expression of pro-inflammatory cytokines in M1 polarized BV-2 microglial cells following LPS treatment. The effects of α-LA on microglial polarization was analyzed through both immunofluorescence and RT-qPCR. BV-2 microglial cells were treated with LPS (1 μg/ml) for 30 min and by indicated concentrations of α-LA at the suggested times. (A, B and D) The cytokine IL-1β, the cell surface marker ICAM1 and ARG1 mRNA expression in the total RNA were assessed through real-time quantitative PCR. (C) BV-2 microglial cells were visualized using an anti-MRC1 antibody and an Alexa Fluor ® 488-linked secondary antibody. Successively, coverslips were detected via fluorescence microscopy. Scale bar = 100 μm.

Article Snippet: In contrast, the antibodies employed for immunofluorescence labeling analysis are the anti-Mannose receptor, goat anti-mouse IgG H&L Alexa Fluor ® 488, and goat anti-rabbit IgG H&L Alexa Fluor ® 488 (Abcam, Milton, Cambridge, UK).

Techniques: Expressing, Immunofluorescence, Quantitative RT-PCR, Marker, Real-time Polymerase Chain Reaction, Fluorescence, Microscopy