Goat anti mouse igg h l alexa fluor 647 antibody

After dissection, the rat brain tissues were immediately fixed in 10% (v/v) neutral formalin solution. Then, the tissues were embedded in paraffin and cut to 5 μm thick. The slides were deparaffinized and rehydrated before immunofluorescence staining using the anti-TLR4 antibody overnight at 4°C. After washing with phosphate-buffered saline (PBS, ph = 7.4), TLR4-positive cell detection was enhanced by using a Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) antibody (Abcam, Cambridge, MA) in the dark. Moreover, the nuclei were stained with 4′,6′-diamidino-2-phenylindole (DAPI, 2 μg/ml). The stained slides were observed on a CRI fluorescence imaging system (Maestro2, CRI, United States) and analyzed using ImageJ software.

Grow cells in 6-well plates containing confocal slides (1 × 10⁴ cells per well). After washing with PBS, cells were fixed with 4% paraformaldehyde for 15 min, and 0.5%Triton X-100 was permeable for 20 min. After the slides were soaked with PBS, 1%bovine serum albumin was blocked for 30 min, each slides were dripped with sufficient amount of diluted Paxillin antibody (Abcam, UK, Cat. no. ab32084) and incubated overnight at 4℃ in a wet box. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody (Abcam, UK, Cat. no. ab150077) was added in the dark and incubated in a wet box at 37℃. Then, cells were incubated overnight at 4℃ in a wet box with diluted PTBP1 antibody (Invitrogen, USA, Cat. no. 32-4800), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) antibody (Abcam, UK, Cat. no. ab150115). Hoechst 33342 (Beyotime Biotechnology, China, Cat. no. C1025) was added to the drops and incubated for 5 min, and the specimens were nucleated. The tablets were sealed with a sealing solution containing an anti-fluorescence quench agent, and the acquired images were observed under a confocal laser scanning microscope (Carl Zeiss, Germany) at 200x magnification.