After behavioral experiments, all animals were euthanized with chloral hydrate (350 mg/kg) during the estrus phase. To measure histology analysis, we performed hematoxylin and eosin (H&E) and Masson's trichrome staining and the clitoral and vaginal tissues were either fixed in 10% buffered formalin or snap frozen in liquid nitrogen. Blood was collected by cardiac puncture. Serum was extracted from the blood samples. To perform enzyme-linked immunosorbent assay (ELISA), tissue homogenates were prepared in ice-cold buffer containing 50 mM Tris and 150 mM NaCl (pH 8.0) and supplemented with 1 mM EDTA, 0.1 mM phenylmethanesulfonyl fluoride, and 1 μg/mL each of aprotinin, leupeptin, and pepstatin (Beyotime Biotech, Shanghai, China) using a homogenizer (OSE-Y50; TIANGEN, Beijing, China), followed by centrifugation at 11,000 g for 15 minutes at 4°C.
Pepstatin
Manufactured by Beyotime
Sourced in China
Pepstatin is a naturally occurring peptide inhibitor of aspartic proteases. It functions by binding to and inhibiting the active site of aspartic proteases, which are a class of enzymes involved in the breakdown of proteins.
Automatically generated - may contain errors
Lab products found in correlation
Leupeptin, by Beyotime (2 mentions)
Ose y50, by Tiangen Biotech (1 mentions)
Aprotinin, by Beyotime (1 mentions)
Mem per plus membrane protein extraction kit, by Thermo Fisher Scientific (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions)
Nb100 105, by Novus Biologicals (1 mentions)
Nb100 122, by Novus Biologicals (1 mentions)
Ab4729, by Abcam (1 mentions)
Chemi xr 5, by Syngene (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Ab1791, by Abcam (1 mentions)
Ab1191, by Abcam (1 mentions)
2 protocols using pepstatin
1
Histological and Biochemical Analyses of Reproductive Tissue
2
Membrane Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested and lysed using RIPA buffer (Beyotime) supplemented with PMSF (Beyotime), leupeptin (Beyotime) and pepstatin (Beyotime). The membrane proteins were extracted by Mem-PER™ Plus Membrane Protein Extraction kit (Thermo Fisher, USA). The protein concentrations of lysate were determined by bicinchoninic acid (BCA) assay. Protein extracvts were subjected to SDS-PAGE analysis and transferred to polyvinylidene difluoride (PVDF) membrane after electrophoresis. The membranes were blocked with 5% BSA buffer followed by antibody hybridization and then visualized in G-BOX gel imaging system (Chemi XR 5, Syngene). Antibodies and dilution ratio in this study include anti-OCT2 (HPA008567, Sigma-Aldrich) at 1:200 dilution, anti-GAPDH (KC-5G4, Kangchen) at 1:5000 dilution, anti-ATP2A2 (AB54032, Sangon Biotech) at 1:5000 dilution, anti-ENT1 (11337-1-AP, proteintech) at 1:1000 dilution, anti-HIF-1α (610958, BD Biosciences), (NB100-105, Novus) at 1:800 dilution, anti-HIF-2α (NB100-122, Novus) at 1:1000 dilution, anti-H3K27ac (ab4729, Abcam) at 1:3000 dilution, anti-H3K18Ac (ab1191, Abcam) at 1:3000 dilution and anti-H3 (ab1791, Abcam) at 1:3000 dilution.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!