Goat anti rabbit hrp secondary antibody

Paraffin-embedded LTs were cut into 5 μm-thick cross-sections and deparaffinized prior to staining using a standard protocol. For immunohistochemical staining, LTs were deparaffinized and rehydrated. Next, the sections were blocked with 3% H₂O₂ in methanol for 15 min to inactivate endogenous peroxidases and then incubated overnight at 4℃ with one of the following primary antibodies: CD68 (No. 28058-1-AP; Proteintech, Wuhan, China). The sections were then incubated for 30 min at room temperature with a goat anti-rabbit HRP secondary antibody (No. PK10006; Proteintech). All sections were examined under an Olympus B × 40 upright light microscope (Olympus, Tokyo, Japan).

Total protein was extracted from tissues or cells using RIPA buffer (Beyotime, China), supplementing with 1 mmol/L PMSF. Then the protein concentration was measured by the BCA Assay Kit (Beyotime, China). 50 μg of proteins were separated by 10% SDS-PAGE and transferred to a PVDF membrane. Proteins were probed with Egr2 primary antibody (Cat # 13491-1-AP, 1:750, Proteintech) and GAPDH (Cat # 10494-1-AP, 1:5000, Proteintech), respectively. The blots were incubated with a goat anti-rabbit HRP secondary antibody (Cat #SA00001-2, 1:5000, Proteintech). Finally, the integrated density of the band was detected using an ECL Detection Reagent (Millipore, MA) and quantified by Image Lab software (Bio-Rad, USA).