Identification of 2E7 mAb Target Protein

For identifying the target protein of mAb 2E7, ARPE-19 cells (10 million) lysed with 0.5% NP40 lysis mix^{26 (link)} were incubated with 2E7 mAb pre-bound to protein G-agarose beads (RepliGen) for 6 h at 4 °C. The recovered proteins were resolved on a SDS-polyacrylamide gel (12.5%) and visualized using Gel Code Blue Reagent (Thermo Fisher). Polypeptides migrating at 60 kDa and 50 kDa were excised and subjected to mass spectrometry proteomic analysis at Bioproximity, Inc (Chantilly, VA)^{31 (link)}.

Free full text: Click here

Stein K.R., Gardner T.J., Hernandez R.E., Kraus T.A., Duty J.A., Ubarretxena-Belandia I., Moran T.M, & Tortorella D. (2019). CD46 facilitates entry and dissemination of human cytomegalovirus. Nature Communications, 10, 2699.

Publication 2019

protein G-agarose beads Gel Code Blue Reagent

Agarose Cells Code blue Mass spectrometry Polyacrylamide gel Polypeptides Protein g Protein target Proteins

Corresponding Organization :

Other organizations : Icahn School of Medicine at Mount Sinai

Top 5 similar protocols

Variable analysis

independent variables

Incubation of ARPE-19 cell lysates with 2E7 mAb pre-bound to protein G-agarose beads

dependent variables

Proteins recovered from the 2E7 mAb immunoprecipitation, as visualized by SDS-polyacrylamide gel electrophoresis and Gel Code Blue Reagent staining
Identification of the target proteins of mAb 2E7 by mass spectrometry proteomic analysis

control variables

ARPE-19 cell lysates (10 million cells)
0.5% NP40 lysis buffer
Incubation time of 6 h at 4 °C
SDS-polyacrylamide gel (12.5%)

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!