TRIM5α Restriction Factor Saturation Assay

Restriction factor saturation assays were performed as previously described [10] (link). Briefly, TE671 cells expressing endogenous human TRIM5alpha were infected with 2-fold serial dilutions of freshly harvested 293T cell supernatants containing LacZ-encoding VLPs. Cultures were incubated for 4–6 hours before adding a fixed amount of GFP encoding N-MLV. After 72 hours, infected cells were harvested and the percentage of GFP positive cells was determined by flow cytometry using a FACS Calibur analyzer (Becton Dickinson).

Free full text: Click here

Wight D.J., Boucherit V.C., Wanaguru M., Elis E., Hirst E.M., Li W., Ehrlich M., Bacharach E, & Bishop K.N. (2014). The N-Terminus of Murine Leukaemia Virus p12 Protein Is Required for Mature Core Stability. PLoS Pathogens, 10(10), e1004474.

Publication 2014

FACS Calibur analyzer

293t cell Assays Dilutions Flow cytometry Human trim5alpha Lacz

Corresponding Organization :

Other organizations : The Francis Crick Institute, Tel Aviv University

Top 5 similar protocols

Variable analysis

independent variables

Serial dilutions of freshly harvested 293T cell supernatants containing LacZ-encoding VLPs

dependent variables

Percentage of GFP positive cells determined by flow cytometry

control variables

TE671 cells expressing endogenous human TRIM5alpha
Fixed amount of GFP encoding N-MLV

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!