Stable Expression of Fluorescent Reporters

We stably expressed a nuclear histone 2B (H2B) marker and KTRs for Akt and ERK as described previously^{13 (link),69 (link)}. We used a PiggyBac transposon (Systems Biosciences, Palo Alto, CA, USA) with a vector containing H2B-mCherry, Akt KTR-mAquamarine, ERK KTR-mCitrine, and a puromycin selection marker, which we refer to as the pHAEP vector. After transposing, we selected positive cells by culturing cells using 5 μg/ml puromycin (ThermoFisher).

Free full text: Click here

Kinnunen P.C., Humphries B.A., Luker G.D., Luker K.E, & Linderman J.J. (2024). Characterizing heterogeneous single-cell dose responses computationally and experimentally using threshold inhibition surfaces and dose-titration assays. NPJ Systems Biology and Applications, 10, 42.

Publication 2024

PiggyBac transposon puromycin

Corresponding Organization :

Other organizations : University of Michigan–Ann Arbor

Top 5 similar protocols

Variable analysis

independent variables

Stable expression of nuclear histone 2B (H2B) marker
Stable expression of KTRs for Akt and ERK

dependent variables

Measurement of Akt and ERK activities using the respective KTRs

control variables

Use of a PiggyBac transposon vector containing H2B-mCherry, Akt KTR-mAquamarine, ERK KTR-mCitrine, and a puromycin selection marker (pHAEP vector)
Culturing of cells using 5 μg/ml puromycin (ThermoFisher) to select for positive cells after transposing

controls

Positive control: Stable expression of nuclear histone 2B (H2B) marker, Akt KTR-mAquamarine, and ERK KTR-mCitrine
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!