Western Blot Analysis of CUL4A

Western blotting was performed using a sodium dodecyl sulfate-polyacrylamide gel electrophoresis system as described previously [12 (link)]. Immunoblotting was performed by incubation at 4 °C with antibodies against CUL4A (1:1000; CST) and β-actin (1:2000, Santa Cruz Biotechnology) overnight. Blots were then washed and incubated with a 1:2000 dilution of horseradish peroxidase (HRP)-conjugated secondary antibody (Jackson, West Grove, PA, USA), followed by three washes with Tris-buffered saline-containing Tween 20. Pierce Enhanced chemiluminescent HRP substrate (Thermo Fisher) was used for detection according to the manufacturer’s instructions.

Free full text: Click here

Huang G.K., Liu T.T., Weng S.W., You H.L., Wei Y.C., Chen C.H., Eng H.L, & Huang W.T. (2017). CUL4A overexpression as an independent adverse prognosticator in intrahepatic cholangiocarcinoma. BMC Cancer, 17, 395.

Publication 2017

β-actin horseradish peroxidase (HRP)-conjugated secondary antibody Pierce Enhanced chemiluminescent HRP substrate

Actin Antibodies Antibody Dilution Horseradish peroxidase Saline Sodium dodecyl sulfate polyacrylamide gel electrophoresis Tween 20

Corresponding Organization :

Other organizations : Kaohsiung Chang Gung Memorial Hospital, Chang Gung University, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung Medical University Chung-Ho Memorial Hospital

Top 5 similar protocols

Protocol cited in 1 other protocol

Variable analysis

independent variables

Antibodies against CUL4A
Antibodies against β-actin

dependent variables

Protein expression levels of CUL4A and β-actin

control variables

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis system
Incubation with antibodies at 4 °C overnight
Washing and incubation with horseradish peroxidase (HRP)-conjugated secondary antibody
Washing with Tris-buffered saline-containing Tween 20
Detection using Pierce Enhanced chemiluminescent HRP substrate

controls

Positive control: Not explicitly mentioned
Negative control: 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!