Whole-cell lysate preparation protocol

For preparing whole-cell lysate, SN12C and SN12L1 cells were washed twice with cold phosphate-buffered saline (PBS), harvested, and lysed with 0.1% sodium dodecyl sulfate (SDS)-added radioimmunoprecipitation assay buffer (Thermo Fisher Scientific, Tokyo, Japan) [49 (link)]. Cell fractions were extracted using a Cell Fractionation Kit (Abcam), according to the manufacturer’s instructions [50 (link)]. Protein assay was performed using a Protein Assay Rapid Kit (Wako Pure Chemical Corporation, Osaka, Japan).

Free full text: Click here

Owari T., Sasaki T., Fujii K., Fujiwara-Tani R., Kishi S., Mori S., Mori T., Goto K., Kawahara I., Nakai Y., Miyake M., Luo Y., Tanaka N., Kondoh M., Fujimoto K, & Kuniyasu H. (2020). Role of Nuclear Claudin-4 in Renal Cell Carcinoma. International Journal of Molecular Sciences, 21(21), 8340.

Publication 2020

radioimmunoprecipitation assay buffer Cell Fractionation Kit Protein Assay Rapid Kit

A protein Assay Buffer Cell Cell fractionation Cold Phosphate Protein Radioimmunoprecipitation assay Saline Sodium dodecyl sulfate

Corresponding Organization : Nara Medical University

Other organizations : Nantong University, Osaka University

Top 5 similar protocols

Variable analysis

independent variables

Cell line (SN12C and SN12L1)

dependent variables

Protein levels in whole-cell lysate
Protein levels in cell fractions

control variables

Phosphate-buffered saline (PBS) for washing cells
Radioimmunoprecipitation assay (RIPA) buffer with 0.1% sodium dodecyl sulfate (SDS) for cell lysis
Cell Fractionation Kit (Abcam) for extracting cell fractions
Protein Assay Rapid Kit (Wako Pure Chemical Corporation) for protein assay

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!