The formalin-fixed and paraffin-embedded sections were deparaffinized and rehydrated. After rinsing with phosphate buffered saline (PBS), the sections were boiled in 10 mM sodium citrate buffer (pH 6.0) for antigen retrieval and incubated in freshly made 3% H2O2 to endogenous peroxidase. After washing three times with PBS, the sections were incubated with PKCε (1:50, SC-214, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or IRS-1 tyr (1:50, SC-17196, Santa Cruz Biotechnology) antibodies at 4°C overnight. The next day, the sections were incubated with the corresponding secondary antibodies and then with diaminobenzidine (DAB) for color development. After counterstaining with hematoxylin, the sections were dehydrated, mounted, and photographed. Image-Pro Plus (Media Cybernetics Co., Rockville, MD, USA) was used for the grayscale scanning and analysis.
Sc 17196
Manufactured by Santa Cruz Biotechnology
Sourced in United States
SC-17196 is a laboratory equipment product from Santa Cruz Biotechnology. It is designed for general laboratory use, but a detailed description of its core function is not available at this time.
Automatically generated - may contain errors
Lab products found in correlation
Image pro plus, by Media Cybernetics (1 mentions)
Sc 214, by Santa Cruz Biotechnology (1 mentions)
Anti actin, by Merck Group (1 mentions)
Anti xiap, by Cell Signaling Technology (1 mentions)
Anti cyclin d1, by Cell Signaling Technology (1 mentions)
Anti p irs 1 2y612, by Santa Cruz Biotechnology (1 mentions)
Tgf β, by R&D Systems (1 mentions)
Mccoy s 5a serum free medium, by Merck Group (1 mentions)
Anti p smad2 s465 467, by Cell Signaling Technology (1 mentions)
Anti irs1, by Cell Signaling Technology (1 mentions)
Anti smad3, by Cell Signaling Technology (1 mentions)
2 protocols using sc 17196
1
Immunohistochemistry for PKCε and IRS-1
2
Colon Cancer Cell Line Culturing and Treatments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human colon cancer cell lines, FET, FET-DNRII, CBS, and CBS-RII, were described previously [32 (link)]. All cells were cultured in McCoy’s 5A serum-free medium (Sigma) supplemented with 10 ng/ml epidermal growth factor (EGF), 20 μg/ml insulin, and 4 μg/ml transferrin [44 (link)]. When cells were subjected to growth factor deprivation stress (GFDS), they were cultured in McCoy’s 5A serum-free medium in the absence of EGF, insulin and transferrin. Cells were maintained at 37°C in a humidified incubator with 5% CO2. TGFβ and the TGFβ RI kinase inhibitor were obtained from R&D Systems and Calbiochem respectively. The following antibodies were used in this study: anti-IRS-1 (1:1000), Cell Signaling #2382, anti-p-IRS-1Y632 (1:600), Santa Cruz sc-17196, anti-p-IRS-1/2Y612 (1:600), Santa Cruz sc-17195-R, anti-Smad3 (1:1000), Cell Signaling #9523, anti-p-Smad3S423/425 (1:1000), Cell Signaling #9520, anti-p-Smad2S465/467 (1:1000), Cell Signaling #3101, anti-XIAP (1:1000), Cell Signaling #14334, anti-Cyclin D1 (1:1000), Cell Signaling #2978 and anti-Actin (1:1000), Sigma #A2066.
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!