Rabbit anti tlr9

Primary antibodies including rabbit anti-cleaved CASP-3 (1:200), goat anti-NLRP3 (1:200, Abcam), rabbit anti- RIPK3 (1:200), rabbit anti-TLR9 (1:200), and mouse anti- NeuN (1:500, Abcam) were used to detect TLR9 expression with neurons. Sections were incubated with DyLight Cy3 anti-rabbit IgG or Alexa488 anti-mouse IgG (1:500, Jackson ImmunoResearch, West Grove, PA, USA), and the nuclei were stained with 4′,6-diamidino-2-phenyl- indole (1:500, Beyotime). Five cortical regions from each rat were randomly selected. The fluorescent signal intensity of the target proteins was detected by a confocal laser scanning microscope (Olympus, Tokyo, Japan).^{[31 (link)]}

After the rats were sacrificed, the cortex was separated on ice, homogenized, and centrifuged. The supernatant was extracted and loading buffer (Beyotime, Shanghai, China) was mixed. The samples were separated and then transferred to membranes (Millipore, Billerica, MA, USA). Primary antibodies, including rabbit anti-TLR9, rabbit anti-CASP-3, mouse anti-RIPK1, rabbit anti-RIPK3 (1:1000, Abcam, Cambridge, MA, USA), rabbit anti-Bcl-2, goat anti-NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3), rabbit anti-CASP-1, rabbit anti- phospho-mixed lineage kinase domain-like protein (MLKL) (Ser345), rabbit anti-MAPKAP kinase 2 (MK2), rabbit anti-phospho-MK2 (Thr334) (1:500, Abcam), rabbit anti-ERK, rabbit anti-phospho-ERK (Thr202/Tyr204), rabbit anti-JNK, and rabbit anti-phospho-JNK (Thr183/Tyr185; 1:1000, Cell Signaling Technology), and mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:5000, ZSGB-BIO, Beijing, China), were incubated overnight. Then, horseradish peroxidase-conjugated anti-goat/rabbit/mouse (1:5000, ZSGB-BIO) IgGs were incubated for 1 h. The blots were visualized using the electrochemiluminescence method (Millipore) and imaged with a ChemiDoc^TM MP imaging system (Bio-Rad, Hercules, CA, USA).^{[31 (link)]}