Four-micron-thick paraffin-embedded colon sections from the AOM/DSS-induced colorectal cancer models were deparaffinized in xylene (Sinopharm) and rehydrated through a graded series of ethanol solutions (Sinopharm). After blocking endogenous peroxidase activity in 3% hydrogen peroxide (ZSGB-BIO) at room temperature for 15 minutes, the tissue sections were treated with 0.01 mol/L citrate buffer (pH 6.0; OriGene) under high pressure in a pressure cooker for 3 minutes to complete antigen retrieval. Blocking was performed with 10% bovine serum (Gibco) for 30 minutes at room temperature, and then the sections were incubated with primary antibodies to CD8a (1:500) and MRP8 (1:1,000) overnight at 4°C (Supplementary Table S2), followed by incubation with 100 μL goat anti-rabbit secondary antibody (ZSGB-BIO, catalog no. PV6001) at room temperature for 30 minutes. Staining was visualized with 3,3′-diaminobenzidine (DAB) (ZSGB-BIO, catalog no. PV8000), and sections were counterstained with hematoxylin, dehydrated, and covered with a coverslip. The slides were scanned using the NanoZoomer digital slice scanner (Hamamatsu). Image analysis and quantitation were performed with ImageJ (NIH, Bethesda, MD).
Citrate buffer
Manufactured by OriGene
Sourced in China
Citrate buffer is a chemical solution used in various laboratory applications. It maintains a specific pH range to create an optimal environment for certain reactions or processes. The buffer helps to control and stabilize the pH level in the solution.
Automatically generated - may contain errors
2 protocols using citrate buffer
1
Immunohistochemical Evaluation of Colon Tumors
2
Immunocytochemistry and Immunohistochemistry Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For ICC, cells grown on coverslips were fixed with cold methanol for 30 min. After treatment with 0.3% endogenous peroxidase inhibitor (Zhongshan Inc., Beijing, China), cells were incubated with goat serum (Zhongshan Inc.) to block non-specific protein binding. The cells on coverslips were then incubated with primary antibodies, biotinylated goat anti-mouse/rabbit IgG (Zhongshan Inc.), and horseradish peroxidase-labeled streptomycin (Zhongshan Inc.). Paraffin-embedded tissue sections were deparaffinized in xylene for IHC. Antigen retrieval was performed by incubation of sections with citrate buffer (OriGene, Wuxi, China). The sections were then incubated with primary antibodies and biotinylated goat anti-rabbit IgG. The signal was detected with a labeled streptavidin-biotin system in the presence of the chromogen 3,3-diaminobenzidine or alkaline phosphatase.
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!