Dear all,

I'm planning to transfect my protein of interest into the adherent cells by using electroporation method. However hardly find a protocol on this method. Does anyone transfect protein into adherent cells by electroporation method before? Could kindly provide me the protocol? Thank you very much.

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Hi Caryn,
People normally transfect plasmid DNA into cells to express their protein of interest. I have heard of protein transfection but my understanding was it was difficult and depended on the properties of the protein. Maybe you could ask the technical support at either Amaxa or BioRad, both companies who make electroporators.

I've also seen use of cell permeable peptides or proteins used, some incorporating a portion of retroviral Gag sequence to get them into the cell.

Ceri

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This is an example "TAT Protein Transduction Domain". They r used to deliver biologically active proteins across membranes. Also used for invivo studies

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My mistake sorry.

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Actually I have tried to transfect my proteins into the adherent cells by using the commercial protein transfection kit, but doesn't work. So, plan to change to other methods like microinjection or electroporation, and I am interested in doing protein electroporation.

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Hi,
There is a new electroporation system available. It electroporates in a pipette tip rather than a cuvette. We've seen really good transfection efficiencies and cell viabilities using it. I've not tried proteins, but the literature says it can be used for those too. We just had a "microporator" on demo from Labtech, and were allowed to borrow it for a week, so I guess you'll be able to as well.
Hope this helps!
Good luck.
James

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We inject a reporter and electroporate it. I am not sure if its the same as proteins though.

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