To explore the effects of PNGL on HMGB expression and IBA1-marked microglia activation in CUMS-induced rats [42 (link),43 (link)], immunofluorescence staining was performed as previously described [7 (link),8 (link),44 (link),45 (link)]. Briefly, after the micro slides were deparaffinized, dewatered, and restored with a citrate-EDTA antigen retrieval solution (P0086, Beyotime, Shanghai, China) for 20 min at 95 ℃, they were cooled down and washed with PBS three times (10 min per time), blocked with 5% goat serum albumin at room temperature for 60 min, and then incubated overnight with an anti-HMGB1 antibody (ab79823, 1:500 in dilution) and IBA1 (ab15690, 1:400 in dilution) at 4 °C. Subsequently, they were incubated with a TRITC-conjugated goat anti-rabbit IgG at a 1:100 dilution (CW0160, CWBIO, Beijing, China) and a Alexa Fluor 488-labeled goat anti-mouse IgG (P0188, Beyotime, Shanghai, China) for 2 h at room temperature, and then counterstained by DAPI (5 μg/mL) for 10 min. Images were observed using fluorescence microscopy (Leica, Germany Q9). The fluorescence intensity was evaluated by the ImageJ 1.44p software.
P0086
Manufactured by Beyotime
Sourced in China
P0086 is a laboratory equipment product. It is designed to perform a core function related to laboratory tasks. The description of its intended use and technical specifications is not available at this time.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using p0086
1
Exploring HMGB1 and Microglia Activation in CUMS-Induced Rats
2
Immunohistochemical Detection of UCP1 in Rat Adipose Tissue
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Rat abdominal subcutaneous adipose tissue slices were immersed in antigen retrieval solution (P0086, Beyotime, Shanghai, China) and boiled at 95°C for 10 min to repair antigen. Ten-minute treatment with 3% H2O2 was then conducted to remove endogenous peroxidases in the slices. Later, the slices were blocked for 30 min using 5% bovine serum albumin (BSA; B928042; MACKLIN, China) at 37°C and incubated with primary antibody for UCP1 (ab234430; Abcam, Cambridge, UK) at 4°C overnight. Afterward, secondary antibody HRP-conjugated goat anti-rabbit IgG (31460, ThermoFisher, Waltham, MA, USA) was used to probe the slices for 1 h in the dark, followed by color development using DAB solution (D8001; Sigma-Aldrich, USA). The slices were counterstained with hematoxylin and observed via the optical microscope under 100× magnification.
3
STING Expression After Subarachnoid Hemorrhage
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At 24 h post-SAH, rats were anesthetized and transcranial perfused with 250 mL PBS (0.1 M, pH 7.4) followed by 500 mL 4% paraformaldehyde. Then, 4- μm-thick coronal brain sections (n = 4/group) were deparaffinized in xylene, rehydrated via an alcohol gradient and washed with PBS (0.01 M, pH 7.4). Immunohistochemical staining was performed based on the commercial kit (SPN-9002, ZSGB-Bio, Beijing, China). Antigen retrieval was performed with citrate-EDTA antigen retrieval solution (P0086, Beyotime, China) in a microwave oven lasting for 25 min. After that, endogenous peroxidase activity was blocked employing 0.3% H2O2 for 10 min and then washed with PBS. After blocked by 5% goat serum for 15 min, slices were incubated overnight at 4 °C with the mouse monoclonal anti-STING antibody (1 : 100, NBP2-24683, Novus). Adjacently, sections were incubated with biotinylated goat antimouse IgG secondary antibody for 15 min and later with HRP-streptavidin reagent. Ultimately, immunoreactivity was visualized using 3,3-diaminobenzidine (Boster), followed by restaining with hematoxylin. Images were obtained via a light microscope (Leica, DM2500, Germany).
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