Protocol detail

Western Blot Analysis of Signaling Proteins

Find Similar Protocols

Total protein was collected from the cells after various treatments. For Western blots, a previously described procedure was applied [28 (link)]. The following primary antibodies were used: p-Jak2 (Y1007/1008), total Jak2, p-Stat3 (Y705), p-Stat3 (S727), and total Stat3 were purchased from Cell Signaling Technology. Antibodies for gp130, gp80, Bmi-1, Oct4 and GAPDH were from Santa Cruz. Mcl-1 and CyclinD1 antibodies were obtained from Abcam. The secondary anti-mouse and anti-rabbit antibodies were purchased from Santa Cruz. All the experiments were performed at least twice and quantification data with statistical analysis was summarized in Figure S19.

Free full text: Click here

Zhao H., Guo Y., Li S., Han R., Ying J., Zhu H., Wang Y., Yin L., Han Y., Sun L., Wang Z., Lin Q., Bi X., Jiao Y., Jia H., Zhao J., Huang Z., Li Z., Zhou J., Song W., Meng K, & Cai J. (2015). A novel anti-cancer agent Icaritin suppresses hepatocellular carcinoma initiation and malignant growth through the IL-6/Jak2/Stat3 pathway. Oncotarget, 6(31), 31927-31943.

Publication 2015

p-Jak2 (Y1007/1008), total Jak2 total Stat3 secondary anti-mouse and anti-rabbit antibodies

Anti antibodies Antibodies Cells Gapdh Gp130 Gp80 Jak2 Mouse Oct4 Protein Rabbit Stat3 Western blots

Top 5 similar protocols

Variable analysis

independent variables

Various treatments

dependent variables

Total protein collected from the cells
Phosphorylation of Jak2 (Y1007/1008)
Total Jak2 levels
Phosphorylation of Stat3 (Y705)
Phosphorylation of Stat3 (S727)
Total Stat3 levels
Gp130 levels
Gp80 levels
Bmi-1 levels
Oct4 levels
Mcl-1 levels
CyclinD1 levels

control variables

GAPDH levels (as a loading control)

controls

Positive controls: Not explicitly mentioned.
Negative controls: Not explicitly mentioned.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!