Delivering defined amounts of purified protein with high precision into living cells

Kristen Skruber; Tracy Ann Read; Eric A. Vitriol

doi:10.1016/j.xpro.2020.100272

Delivering defined amounts of purified protein with high precision into living cells

Kristen Skruber, Tracy Ann Read, Eric A. Vitriol

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

Here, we detail a protocol using electroporation to precisely deliver defined amounts of purified protein into CAD cells. This method allows one million cells to be electroporated with protein simultaneously, with high delivery efficiency and low cell death. Further, by circumventing the normal biosynthetic pathway, proteins can be studied without the complication of post-translational modifications and before a transcriptional response can be initiated. This protocol will be useful for any researcher who is interested in protein concentration-dependent cellular phenotypes. For complete details on the use and execution of this protocol, please refer to Skruber et al. (2020).

Original language	English (US)
Article number	100272
Journal	STAR Protocols
Volume	2
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2020.100272
State	Published - Mar 19 2021
Externally published	Yes

Keywords

Cell biology
Cell culture
Cell-based assays
Microscopy
Molecular/chemical probes
Protein expression and purification

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Neuroscience(all)
Immunology and Microbiology(all)
Medicine(all)

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10.1016/j.xpro.2020.100272

Cite this

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title = "Delivering defined amounts of purified protein with high precision into living cells",

abstract = "Here, we detail a protocol using electroporation to precisely deliver defined amounts of purified protein into CAD cells. This method allows one million cells to be electroporated with protein simultaneously, with high delivery efficiency and low cell death. Further, by circumventing the normal biosynthetic pathway, proteins can be studied without the complication of post-translational modifications and before a transcriptional response can be initiated. This protocol will be useful for any researcher who is interested in protein concentration-dependent cellular phenotypes. For complete details on the use and execution of this protocol, please refer to Skruber et al. (2020).",

keywords = "Cell biology, Cell culture, Cell-based assays, Microscopy, Molecular/chemical probes, Protein expression and purification",

author = "Kristen Skruber and Read, {Tracy Ann} and Vitriol, {Eric A.}",

note = "Funding Information: This project was supported by a National Institutes of Health (NIH) Pathway to Independence Award (R00 NS087104) and an NIH Maximizing Investigators{\textquoteright} Research Award for Early Stage Investigators (R35 GM137959) to E.A.V. Conceptualization, K.S. and E.A.V.; Methodology, K.S. and E.A.V.; Formal Analysis, K.S. and E.A.V.; Investigation, K.S. and E.A.V.; Resources, K.S. and E.A.V.; Writing, K.S. T.A.R. and E.A.V.; Visualization, K.S. and E.A.V.; Project Administration, E.A.V.; Supervision, E.A.V.; Funding Acquisition, E.A.V. The authors declare no competing interests. Funding Information: This project was supported by a National Institutes of Health (NIH) Pathway to Independence Award ( R00 NS087104 ) and an NIH Maximizing Investigators{\textquoteright} Research Award for Early Stage Investigators ( R35 GM137959 ) to E.A.V. Publisher Copyright: {\textcopyright} 2020 The Author(s)",

year = "2021",

month = mar,

day = "19",

doi = "10.1016/j.xpro.2020.100272",

language = "English (US)",

volume = "2",

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AU - Read, Tracy Ann

AU - Vitriol, Eric A.

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N2 - Here, we detail a protocol using electroporation to precisely deliver defined amounts of purified protein into CAD cells. This method allows one million cells to be electroporated with protein simultaneously, with high delivery efficiency and low cell death. Further, by circumventing the normal biosynthetic pathway, proteins can be studied without the complication of post-translational modifications and before a transcriptional response can be initiated. This protocol will be useful for any researcher who is interested in protein concentration-dependent cellular phenotypes. For complete details on the use and execution of this protocol, please refer to Skruber et al. (2020).

AB - Here, we detail a protocol using electroporation to precisely deliver defined amounts of purified protein into CAD cells. This method allows one million cells to be electroporated with protein simultaneously, with high delivery efficiency and low cell death. Further, by circumventing the normal biosynthetic pathway, proteins can be studied without the complication of post-translational modifications and before a transcriptional response can be initiated. This protocol will be useful for any researcher who is interested in protein concentration-dependent cellular phenotypes. For complete details on the use and execution of this protocol, please refer to Skruber et al. (2020).

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Delivering defined amounts of purified protein with high precision into living cells

Abstract

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ASJC Scopus subject areas

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