Protein samples were extracted from 0.5 g of gastro tissue using a protein extraction kit (Beyotime Biotechnology, China); these samples were then used for western blot analysis, which was performed according to the manufacturer’s instructions. A dilution ratio of 1:1000 was used for all the primary antibodies, including rabbit anti-NLPP3 polyclonal antibody (19771–1-AP, Proteintech), rabbit anti-TLR4 polyclonal antibody (A5258, Abclonal), rabbit anti-p65 monoclonal antibody (CST 8242T, CST, Danvers, MA, USA), rabbit anti-GSDMD monoclonal antibody (CST 69469S, CST), rabbit anti-Caspase 1 monoclonal antibody (CST 3866T, CST, Danvers), mouse anti-iκBα monoclonal antibody (CST 4814T, CST), rabbit anti-Myd88 polyclonal antibody (A0980, Abclonal), rabbit anti-IL-1β polyclonal antibody (A16288, Abclonal), rabbit anti-IL-18 polyclonal antibody (A1115, Abclonal), rabbit anti-ZO-1 monoclonal antibody (CST 13,663, CST), rabbit anti-Occludin monoclonal antibody (CST 91,131, CST), rabbit anti-RORγ polyclonal antibody (A10240 Abclonal), rabbit anti-Foxp3 monoclonal antibody (CST12632; CST), and mouse anti-β-actin monoclonal antibody (CST4970; CST). A dilution ratio of 1:2000 was used for all the secondary antibodies, including anti-rabbit IgG, HRP-linked antibody (CST7074, CST). Enhanced chemiluminescence was used for detection.
A0980
Manufactured by ABclonal
Sourced in United States
A0980 is a laboratory equipment product manufactured by ABclonal. It is designed for general laboratory use. The core function of this product is to provide a controlled environment for various scientific experiments and procedures.
Automatically generated - may contain errors
2 protocols using a0980
1
Western Blot Analysis of Gastro Tissue Proteins
2
Immunohistochemical Analysis of Bone and Adipose Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Femora, tibiae, and BAT dissected from the mice were fixed using 4% paraformaldehyde in phosphate-buffered saline (PBS) at 4°C for 24 h. Brain dissected from the mice which were transcardially perfused with saline followed by 4% paraformaldehyde (PFA) were post-fixed in 4% PFA at 4°C for 24 h. Femora and tibiae decalcified in 15% EDTA (pH 7.4) at 4°C for 14 days. The tissues were embedded in paraffin, and 5-μm sagittal-oriented of femora, tibiae, and BAT, or coronal-oriented of brain sections were prepared for histological analyses. The sections were incubated with primary antibodies against perilipin 1 (PLIN1, #9349,1:150,CST), P-P65-S276 (AP0123, 1:100, ABclonal, Woburn, MA, USA), Rheb (15924-1-AP, 1:100, Proteintech), or Myd88 (A0980, 1:50, ABclonal) and labeled with secondary antibodies for 1 h in the dark. After labeling, cells were incubated with DAPI for 5 min. Images were acquired with an Olympus 200 M microscope (Olympus, Tokyo, Japan). The numbers of positively-stained cells in the whole medullary space or bone trabecula per femur or tibia in three sequential sections per mouse in each group were counted using Image Pro Plus software.
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