Abstract
The on-column capture of a specific protein using magnetic beads was applied to SDS-CGE. In a preliminary study, an immunological reaction in the presence of SDS, using a batch method, was attempted. Carbonic anhydrase (CA), α-lactalbumin (LA), and HSA were denatured by heating in the presence of SDS and 2-mercaptoethanol, and then reacted with anti-CA that had been immobilized on magnetic beads. Not only native CA, but also the denatured CA reacted with anti-CA, even in the presence of SDS. Therefore, the on-column capture of denatured CA separated by SDS-CGE was attempted using a two-point detection technique. A mixture of proteins, containing LA, CA, and HSA, were separated by SDS-CGE according to their Mr. The CA was then specifically captured on anti-CA-immobilized magnetic beads, which were packed between two detection windows in the capillary column, during the electrophoresis. The results show that the technique leads to information similar to that obtained by Western blotting, i.e., a protein can be identified by its Mr and reaction with its antibody.
| Original language | English |
|---|---|
| Pages (from-to) | 2262-2266 |
| Number of pages | 5 |
| Journal | Electrophoresis |
| Volume | 28 |
| Issue number | 13 |
| DOIs | |
| Publication status | Published - Jul 2007 |
| Externally published | Yes |
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Keywords
- CGE
- Immunoaffinity
- Magnetic beads
- Protein
ASJC Scopus subject areas
- Clinical Biochemistry
Cite this
On-column capture of a specific protein separated by SDS-CGE using an immunological reaction on magnetic beads. / Kaneta, Takashi; Takahashi, Masanori; Imasaka, Totaro.
In: Electrophoresis, Vol. 28, No. 13, 07.2007, p. 2262-2266.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - On-column capture of a specific protein separated by SDS-CGE using an immunological reaction on magnetic beads
AU - Kaneta, Takashi
AU - Takahashi, Masanori
AU - Imasaka, Totaro
PY - 2007/7
Y1 - 2007/7
N2 - The on-column capture of a specific protein using magnetic beads was applied to SDS-CGE. In a preliminary study, an immunological reaction in the presence of SDS, using a batch method, was attempted. Carbonic anhydrase (CA), α-lactalbumin (LA), and HSA were denatured by heating in the presence of SDS and 2-mercaptoethanol, and then reacted with anti-CA that had been immobilized on magnetic beads. Not only native CA, but also the denatured CA reacted with anti-CA, even in the presence of SDS. Therefore, the on-column capture of denatured CA separated by SDS-CGE was attempted using a two-point detection technique. A mixture of proteins, containing LA, CA, and HSA, were separated by SDS-CGE according to their Mr. The CA was then specifically captured on anti-CA-immobilized magnetic beads, which were packed between two detection windows in the capillary column, during the electrophoresis. The results show that the technique leads to information similar to that obtained by Western blotting, i.e., a protein can be identified by its Mr and reaction with its antibody.
AB - The on-column capture of a specific protein using magnetic beads was applied to SDS-CGE. In a preliminary study, an immunological reaction in the presence of SDS, using a batch method, was attempted. Carbonic anhydrase (CA), α-lactalbumin (LA), and HSA were denatured by heating in the presence of SDS and 2-mercaptoethanol, and then reacted with anti-CA that had been immobilized on magnetic beads. Not only native CA, but also the denatured CA reacted with anti-CA, even in the presence of SDS. Therefore, the on-column capture of denatured CA separated by SDS-CGE was attempted using a two-point detection technique. A mixture of proteins, containing LA, CA, and HSA, were separated by SDS-CGE according to their Mr. The CA was then specifically captured on anti-CA-immobilized magnetic beads, which were packed between two detection windows in the capillary column, during the electrophoresis. The results show that the technique leads to information similar to that obtained by Western blotting, i.e., a protein can be identified by its Mr and reaction with its antibody.
KW - CGE
KW - Immunoaffinity
KW - Magnetic beads
KW - Protein
UR - http://www.scopus.com/inward/record.url?scp=34447518799&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34447518799&partnerID=8YFLogxK
U2 - 10.1002/elps.200600726
DO - 10.1002/elps.200600726
M3 - Article
C2 - 17538922
AN - SCOPUS:34447518799
VL - 28
SP - 2262
EP - 2266
JO - Electrophoresis
JF - Electrophoresis
SN - 0173-0835
IS - 13
ER -