A regularly spaced and self-revealing protein ladder for anti-tag Western blot analysis

Chien Han Kao; Chiu Min Cheng; Kuo Hsiang Chuang; Chih Hung Chuang; Shey Cherng Tzou; Ta Chun Cheng; Yuan Chin Hsieh; Kuang Wen Liao; Yun Ming Wang; Long Sen Chang; Steve R. Roffler; Fang Ming Chen; Tian Lu Cheng

doi:10.1016/j.ab.2012.08.001

A regularly spaced and self-revealing protein ladder for anti-tag Western blot analysis

Chien Han Kao, Chiu Min Cheng, Kuo Hsiang Chuang, Chih Hung Chuang, Shey Cherng Tzou, Ta Chun Cheng, Yuan Chin Hsieh, Kuang Wen Liao, Yun Ming Wang, Long Sen Chang, Steve R. Roffler, Fang Ming Chen, Tian Lu Cheng

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

5 Citations (Scopus)

Abstract

We designed a protein ladder (hereafter referred to as "Mega- tag") that contains 14 of the most commonly used epitope tags fused to molecular weight markers. The Mega-tag ladder can be simultaneously visualized when anti-tag antibodies are used to detect epitope-tagged recombinant proteins in Western blots. The logarithm of molecular weights and relative mobility of the Mega-tag protein ladder are highly correlated (R²= 0.997 ± 0.00232), indicating that the dye-free Mega-tag protein ladder is accurate. It can also serve as a positive control for anti-epitope tag immunoblots. The Mega-tag protein ladder should provide a convenient and precise tool for Western blot analysis.

Original language	English
Pages (from-to)	1-3
Number of pages	3
Journal	Analytical Biochemistry
Volume	431
Issue number	1
DOIs	https://doi.org/10.1016/j.ab.2012.08.001
Publication status	Published - Dec 1 2012

Keywords

14-Epitope tag protein ladder
Anti-epitope tag antibodies
Epitope tags
Protein molecular estimation
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis
Western blot analysis

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.ab.2012.08.001

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title = "A regularly spaced and self-revealing protein ladder for anti-tag Western blot analysis",

abstract = "We designed a protein ladder (hereafter referred to as {"}Mega- tag{"}) that contains 14 of the most commonly used epitope tags fused to molecular weight markers. The Mega-tag ladder can be simultaneously visualized when anti-tag antibodies are used to detect epitope-tagged recombinant proteins in Western blots. The logarithm of molecular weights and relative mobility of the Mega-tag protein ladder are highly correlated (R2= 0.997 ± 0.00232), indicating that the dye-free Mega-tag protein ladder is accurate. It can also serve as a positive control for anti-epitope tag immunoblots. The Mega-tag protein ladder should provide a convenient and precise tool for Western blot analysis.",

keywords = "14-Epitope tag protein ladder, Anti-epitope tag antibodies, Epitope tags, Protein molecular estimation, Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot analysis",

author = "Kao, {Chien Han} and Cheng, {Chiu Min} and Chuang, {Kuo Hsiang} and Chuang, {Chih Hung} and Tzou, {Shey Cherng} and Cheng, {Ta Chun} and Hsieh, {Yuan Chin} and Liao, {Kuang Wen} and Wang, {Yun Ming} and Chang, {Long Sen} and Roffler, {Steve R.} and Chen, {Fang Ming} and Cheng, {Tian Lu}",

note = "Funding Information: This work was supported by grants from the National Research Program for Biopharmaceuticals, National Science Council, Taipei, Taiwan (NSC 100-2325-B-037-001 and 100-2321-B-037-006), the Department of Health, Executive Yuan, Taiwan (DOH100-TD-N-111-010 and DOH100-TD-C-111-002), and the Grant of Biosignature in Colorectal Cancers, Academia Sinica, Taiwan, and Kaohsiung Medical University Research Foundation (KMUER014 and KMUER012). ",

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AU - Kao, Chien Han

AU - Cheng, Chiu Min

AU - Chuang, Kuo Hsiang

AU - Chuang, Chih Hung

AU - Tzou, Shey Cherng

AU - Cheng, Ta Chun

AU - Hsieh, Yuan Chin

AU - Liao, Kuang Wen

AU - Wang, Yun Ming

AU - Chang, Long Sen

AU - Roffler, Steve R.

AU - Chen, Fang Ming

AU - Cheng, Tian Lu

N1 - Funding Information: This work was supported by grants from the National Research Program for Biopharmaceuticals, National Science Council, Taipei, Taiwan (NSC 100-2325-B-037-001 and 100-2321-B-037-006), the Department of Health, Executive Yuan, Taiwan (DOH100-TD-N-111-010 and DOH100-TD-C-111-002), and the Grant of Biosignature in Colorectal Cancers, Academia Sinica, Taiwan, and Kaohsiung Medical University Research Foundation (KMUER014 and KMUER012).

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N2 - We designed a protein ladder (hereafter referred to as "Mega- tag") that contains 14 of the most commonly used epitope tags fused to molecular weight markers. The Mega-tag ladder can be simultaneously visualized when anti-tag antibodies are used to detect epitope-tagged recombinant proteins in Western blots. The logarithm of molecular weights and relative mobility of the Mega-tag protein ladder are highly correlated (R2= 0.997 ± 0.00232), indicating that the dye-free Mega-tag protein ladder is accurate. It can also serve as a positive control for anti-epitope tag immunoblots. The Mega-tag protein ladder should provide a convenient and precise tool for Western blot analysis.

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A regularly spaced and self-revealing protein ladder for anti-tag Western blot analysis

Abstract

Keywords

ASJC Scopus subject areas

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