Immunofluorescence experiments were performed on rat spinal cord sections and rat astrocyte CTX‐TNA2 cell line. Frozen sections of rat spinal cord and slides of CTX‐TNA2 cells were prepared and blocked using QuickBlock Immunostaining block solution (Beyotime). After incubated with primary antibodies: anti‐NeuN (Novus Biologicals, NB1‐92693), anti‐IBA1 (Wako, 011–27991), anti‐GFAP (CST, 3670S), anti‐NFAT5 (Abcam, ab3446), anti‐AURKB (Abcam, ab2254), or anti‐AQP4 (Abcam, ab2254); the corresponding secondary fluorescent antibodies were used to release the signals. The slides were mounted with the permanent mounting medium and imaged under the Leica DMI4000 fluorescence microscope with the DFC365FX camera (Leica).
Quickblock immunostaining block solution
Manufactured by Beyotime
QuickBlock Immunostaining block solution is a ready-to-use blocking agent designed for immunostaining procedures. It is formulated to reduce non-specific background staining and improve signal-to-noise ratio in immunohistochemistry and immunocytochemistry applications.
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2 protocols using quickblock immunostaining block solution
1
Immunofluorescence Analysis of Rat Spinal Cord and Astrocytes
2
Immunostaining of LAMP2 in Cells
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For IF, cells were plated into glass dishes. After fixation with 4% paraformaldehyde (PFA) for 15 min, cells were permeabilized with 0.5% Triton X-100 in phosphate-buffered saline (PBS) for 15 min. After blocking with QuickBlock™ Immunostaining Block Solution (Beyotime) at room temperature for 1 h, cells were incubated with the mouse anti-LAMP2 antibody (1:200 Dilution, Proteintech) at 4 °C overnight, followed by the secondary antibody Alexa Fluor 647-conjugated goat anti-mouse IgG antibody (1:500 dilution, Cell Signaling Technology, Inc.) incubation at room temperature for 1 h. DAPI (Beyotime) was used for the stain of the nucleus. Cells were photographed under the BioTek Cytation C10 Confocal Image Reader (Agilent technologies, City, State) at ×200 or ×400 magnification.
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