Mouse secondary antibody

Western blotting: TIMELESS (Abcam, ab109512, 1/50000), β-actin (Abcam, ab49900, HRP-linked,1/50000), c-Myc (Abcam, ab32072, 1/1000) and PD-L1 (Cell Signaling Technology, #13684, 1/1000), Rabbit secondary antibody (Cell Signaling Technology, #7074, 1/2000).
Immunohistochemistry: TIMELESS (Abcam, ab72458, 1/200), CD8a (Servicebio, GB13429, 1/200), PD-L1 (Servicebio, GB11339A, 1/500), Ki-67 (Servicebio, GB111141, 1/500), Mouse secondary antibody (Servicebio, GB23301, 1/200), Rabbit secondary antibody (Servicebio, GB23303, 1/200).
Immunofluorescence: TIMELESS (Abcam, ab109512, 1/100), c-Myc (Abcam, ab32072, 1/100), CD8a (Servicebio, GB13429, 1/5000), Granzyme B (Abcam, ab255598, 1/2000), active caspase 3 (Servicebio, GB11532, 1/500), Hoechst 33342 (Beyotime, C1027, 1/100), CY3-TSA (Apex Bio, K1051, 1/200), Donkey Anti-Rabbit IgG H&L Alexa Fluor^® 647 (Abcam, ab150075, 1/200).
Flow cytometry: PD-L1 (Cell Signaling Technology, #13684, 1/200), CD8a-PerCP/Cyanine5.5 (Clone 53-6.7, Biolegend 1007341/100), CD45-AF700 (Biolegend 103128, 1/80), Fixable Viability Dye eFluor™ 450 (Invitrogen, 1/500).

Trigeminal ganglion slices with a thickness of 4 μm were then prepared. Sections were dewaxed in xylene. Then the alcohol grade was lowered for rehydration. Sections were incubated with 0.3% Triton X-100 for 15 min, blocked with 5% BSA for 60 min at room temperature, and then incubated with primary antibody overnight at 4°C. PBS was washed three times after each operation. The primary antibody was diluted as follows: CGRP (1:100; Santa Cruz), Iba-1 (1:500; woko), PFKP (1:200; servicebio), GFAP (1:200; GeneTex), NeuN (1:500; proteintech). After washing with PBS, sections were incubated with mouse secondary antibody (488 nm, green; servicebio). Anti-mouse (488 nm, green; servicebio), anti-rabbit (CY3, red; servicebio) and anti-goat (FITC, green; servicebio) were used. DAPI (servicebio) was incubated for 10 min for staining nuclei. Capture images under the objective of an upright microscope.