Small intestine organoids were grown in 300ul of Matrigel in 1 well each of a 6-well dish containing 3 mls of growth media for 4 days post-passage, then treated with 125ng/ml Adriamycin for 4 hrs to induce p53 expression. Organoids were then recovered from the Matrigel using several rinses with cold PBS. Organoid pellets were lysed with Lamelli buffer. Antibodies used for Western blot were: anti-APC (1:400, FE9 clone, Millipore #MABC202), anti-non-phosphorylated β-catenin (1:1000, #8814, Cell Signaling Technology), anti-p53 (1:500, #NCL-p53-505, Novocastra) and anti-actin-HRP (1:10,000, #ab49900, Abcam).
Anti non phosphorylated β catenin
Manufactured by Cell Signaling Technology
Anti-non-phosphorylated β-catenin is a laboratory reagent used to detect the non-phosphorylated form of the β-catenin protein. β-catenin is a key component of the Wnt signaling pathway and plays a role in cell-cell adhesion and gene transcription. This antibody specifically recognizes the non-phosphorylated form of β-catenin.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using anti non phosphorylated β catenin
1
Adriamycin-Induced p53 Expression in Small Intestine Organoids
2
Quantifying Phosphorylated LRP6 in Mouse Retina
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis detected the phosphorylated level of LRP6. Mouse retina was dissected and homogenized for protein extraction. Equal amounts (50 μg) of total protein from each sample were resolved by SDS polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto a nitrocellulose membrane. The membrane was blocked with 5 % nonfat milk and separately blotted with primary antibodies (anti-Non-phosphorylated β-Catenin, Cell Signaling Technology; anti-phosphorylated LRP6, Cell Signaling Technology; and anti-LRP6, self-made). After thorough washes, the membrane was incubated with the peroxidase-conjugated horse anti-rabbit or horse anti-mouse antibody (Vector Lab). The signal was developed with the enhanced chemiluminescence (ECL) system (Pierce, Rockford, IL, USA). Membranes were stripped and re-blotted with anti-β-actin (Sigma-Aldrich, St. Louis, MO, USA) as the loading control. Densitometry of the signal bands on digital images was quantified using FluorChem Q software (ProteinSimple, Santa Clara, CA, USA).
3
Adriamycin-Induced p53 Expression in Small Intestine Organoids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Small intestine organoids were grown in 300ul of Matrigel in 1 well each of a 6-well dish containing 3 mls of growth media for 4 days post-passage, then treated with 125ng/ml Adriamycin for 4 hrs to induce p53 expression. Organoids were then recovered from the Matrigel using several rinses with cold PBS. Organoid pellets were lysed with Lamelli buffer. Antibodies used for Western blot were: anti-APC (1:400, FE9 clone, Millipore #MABC202), anti-non-phosphorylated β-catenin (1:1000, #8814, Cell Signaling Technology), anti-p53 (1:500, #NCL-p53-505, Novocastra) and anti-actin-HRP (1:10,000, #ab49900, Abcam).
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!