Publication

Improved Native Isolation of Endogenous Protein A-Tagged Protein Complexes

LaCava, J., Chandramouli, N., Jiang, H. & Rout, M. P., Apr-2013, (Accepted/In press) In : Biotechniques. 54, 4, p. 213-216 4 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

LaCava, J., Chandramouli, N., Jiang, H., & Rout, M. P. (Accepted/In press). Improved Native Isolation of Endogenous Protein A-Tagged Protein Complexes. Biotechniques, 54(4), 213-216. https://doi.org/10.2144/000114012

Author

LaCava, John ; Chandramouli, Nagarajan ; Jiang, Hua ; Rout, Michael P. / Improved Native Isolation of Endogenous Protein A-Tagged Protein Complexes. In: Biotechniques. 2013 ; Vol. 54, No. 4. pp. 213-216.

Harvard

LaCava, J, Chandramouli, N, Jiang, H & Rout, MP 2013, 'Improved Native Isolation of Endogenous Protein A-Tagged Protein Complexes', Biotechniques, vol. 54, no. 4, pp. 213-216. https://doi.org/10.2144/000114012

Standard

Improved Native Isolation of Endogenous Protein A-Tagged Protein Complexes. / LaCava, John; Chandramouli, Nagarajan; Jiang, Hua; Rout, Michael P.

In: Biotechniques, Vol. 54, No. 4, 04.2013, p. 213-216.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

LaCava J, Chandramouli N, Jiang H, Rout MP. Improved Native Isolation of Endogenous Protein A-Tagged Protein Complexes. Biotechniques. 2013 Apr;54(4):213-216. https://doi.org/10.2144/000114012


BibTeX

@article{2dfcc9bf30504bc396f003f8b07faa36,
title = "Improved Native Isolation of Endogenous Protein A-Tagged Protein Complexes",
abstract = "Here we report a modified peptide reagent useful for the rapid, native elution of protein complexes containing a Protein-A-tagged component. We tested this reagent for the elution of tagged endogenous protein complexes from yeast (Nup53p/Nup170p dimer; Nup1p/Kap95p/Kap60p trimer; pentameric GINS complex) and bacteria (RNAPII holoenzyme). The majority of the affinity-isolated material is released within 15 minutes under mild conditions, and the elution reagent itself is readily depleted from the elution mixture by simple spin column gel filtration. This reagent is ideal for eluting protein complexes after Protein A / IgG affinity isolation when protease cleavage is not possible or not desirable and facile depletion of the elution reagent is needed.",
keywords = "AFFINITY PURIFICATION, BINDING, IGG, DOMAINS, POLYMERASE, RESOLUTION, ELUTION",
author = "John LaCava and Nagarajan Chandramouli and Hua Jiang and Rout, {Michael P.}",
year = "2013",
month = "4",
doi = "10.2144/000114012",
language = "English",
volume = "54",
pages = "213--216",
journal = "Biotechniques",
issn = "0736-6205",
publisher = "FUTURE SCI LTD",
number = "4",

}

RIS

TY - JOUR

T1 - Improved Native Isolation of Endogenous Protein A-Tagged Protein Complexes

AU - LaCava, John

AU - Chandramouli, Nagarajan

AU - Jiang, Hua

AU - Rout, Michael P.

PY - 2013/4

Y1 - 2013/4

N2 - Here we report a modified peptide reagent useful for the rapid, native elution of protein complexes containing a Protein-A-tagged component. We tested this reagent for the elution of tagged endogenous protein complexes from yeast (Nup53p/Nup170p dimer; Nup1p/Kap95p/Kap60p trimer; pentameric GINS complex) and bacteria (RNAPII holoenzyme). The majority of the affinity-isolated material is released within 15 minutes under mild conditions, and the elution reagent itself is readily depleted from the elution mixture by simple spin column gel filtration. This reagent is ideal for eluting protein complexes after Protein A / IgG affinity isolation when protease cleavage is not possible or not desirable and facile depletion of the elution reagent is needed.

AB - Here we report a modified peptide reagent useful for the rapid, native elution of protein complexes containing a Protein-A-tagged component. We tested this reagent for the elution of tagged endogenous protein complexes from yeast (Nup53p/Nup170p dimer; Nup1p/Kap95p/Kap60p trimer; pentameric GINS complex) and bacteria (RNAPII holoenzyme). The majority of the affinity-isolated material is released within 15 minutes under mild conditions, and the elution reagent itself is readily depleted from the elution mixture by simple spin column gel filtration. This reagent is ideal for eluting protein complexes after Protein A / IgG affinity isolation when protease cleavage is not possible or not desirable and facile depletion of the elution reagent is needed.

KW - AFFINITY PURIFICATION

KW - BINDING

KW - IGG

KW - DOMAINS

KW - POLYMERASE

KW - RESOLUTION

KW - ELUTION

U2 - 10.2144/000114012

DO - 10.2144/000114012

M3 - Article

VL - 54

SP - 213

EP - 216

JO - Biotechniques

JF - Biotechniques

SN - 0736-6205

IS - 4

ER -

ID: 117138044