2024-03-29T00:00:04Z
https://nagoya.repo.nii.ac.jp/oai
oai:nagoya.repo.nii.ac.jp:00007076
2023-01-16T03:52:32Z
320:321:322
A New Isoelectric Focusing System for Fast Two-Dimensional Gel Electrophoresis Using a Low-Concentration Polyacrylamide Gel Supported by a Loose Multifilament String
LI, Jinxiang
OGASAWARA, Ayaka
ODAKE, Tamao
UMEMURA, Tomonari
TSUNODA, Kin-ichi
open access
A new isoelectric focusing (IEF) system for two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) has been proposed. In this system, a super-soft and tough IEF gel was achieved by casting polyacrylamide gel down to 2.0% T using a loose multifilament string (LMS) of nylon as a gel support. The IEF apparatus for the LMS-gel, fabricated from acrylic boards, had a cooling water chamber, and eliminated the need of electrode solutions by directly connecting the two ends of individual gels to platinum electrodes. The carrier ampholyte-generated pH gradients using the new IEF system was stable over a long duration of time and a wide range of voltages, and the IEF time became shorter using a 2.0% T gel than using a 4.0% T gel. Also, the LMS-gels prepared in different runs exhibited excellent reproducibility. The new IEF system was applied to 2-D PAGE of a chicken skeletal muscle extract, and it was found that the protein loading capacity, protein entry into the LMS-gels, and protein transfer efficiency from the first-dimensional to the second-dimensional gels were significantly improved by using a low-concentration (2.5% T) gel. Also, proteins of high molecular weight of more than 200 kDa were observed in the 2-D maps, and therefore the new IEF system has a very good potential to be applied for fast 2-D PAGE of high molecular-weight proteins.
日本分析化学会
2005-03
eng
journal article
VoR
http://hdl.handle.net/2237/8750
https://nagoya.repo.nii.ac.jp/records/7076
https://doi.org/10.2116/analsci.20.1673
0910-6340
Analytical Sciences
20
12
1673
1679
https://nagoya.repo.nii.ac.jp/record/7076/files/v20_n12_1673-1679.pdf
application/pdf
819.0 kB
2018-02-19