Amaxa nucleofection protocol

Manufactured by Lonza

The Amaxa nucleofection protocol is a laboratory technique used for the efficient delivery of nucleic acids, such as DNA or RNA, into cells. It utilizes a specialized electroporation-based method to facilitate the direct transfer of these molecules into the cell nucleus. The core function of this protocol is to enable the introduction of genetic material into a wide range of cell types, including difficult-to-transfect cells, for various research and experimental applications.

Automatically generated - may contain errors

Lab products found in correlation

Topflash reporter construct, by Merck Group (2 mentions) Lipofectamine, by Thermo Fisher Scientific (1 mentions) Pierce bicinchoninic acid bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Nucleofection (130 citations) Amaxa nucleofection (40 citations) Nucleofection protocol (4 citations)

2 protocols using amaxa nucleofection protocol

Measuring β-Catenin Signaling Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols

To measure β-catenin-dependent signaling activity, 5×10⁶ PBMCs cells were transfected with 10 μg TOPflash reporter construct (Millipore, Billerica, MA) or FOPflash and renilla plasmid using the Amaxa nucleofection protocol (Amaxa, Gaithersburg, MD), as recommended by the manufacturer. PDA and L-Wnt3a cells were transfected with 3 μg TOPflash reporter construct or FOPflash and renilla plasmid using Lipofectamine (Life Technologies Invitrogen) according to manufacturer’s instructions. Three days later, construct reporter activity was performed using a dual-luciferase reporter assay using 10 to 20 μl of lysate (Promega, Madison, WI). Briefly, the Relative Light Units (RLUs) were normalized to a Renilla luciferase control or μg/ml of protein as indicated. The total protein concentration was measured using a Pierce bicinchoninic acid (BCA) protein assay kit (Thermo Scientific, Waltham, MA).

Richards M.H., Seaton M.S., Wallace J, & Al-Harthi L. (2014). Porcupine Is Not Required for the Production of the Majority of Wnts from Primary Human Astrocytes and CD8+ T Cells. PLoS ONE, 9(3), e92159.

+ Open protocol

+ Expand

Measuring β-catenin Signaling Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols

To measure β-catenin-dependent signaling activity, 5 ×10⁶ PBMCs cells cultured in cRPMI or cRPMI and ACM or cRPMI and Wnt-depleted ACM were transfected with 10µg TOPflash reporter construct (Millipore, Billerica, MA) or FOPflash using the Amaxa nucleofection protocol (Amaxa, Gaithersburg, MD), as recommended by the manufacturer. Three days later, construct reporter activity was performed using a dual-luciferase reporter assay using 10 to 20 µl of lysate (Promega, Madison, WI). The total protein concentration was measured using a Pierce BCA Protein Assay Kit (Thermo Scientific, Waltham, MA).

Richards M.H., Narasipura S.D., Seaton M.S., Lutgen V, & Al-Harthi L. (2015). Migration of CD8+ T cells into the CNS gives rise to highly potent anti-HIV CD4dimCD8bright T cells in a Wnt signaling -dependent manner. Journal of immunology (Baltimore, Md. : 1950), 196(1), 317-327.

+ Open protocol

+ Expand

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!