The measurement of pgam-b using elisa and changes of serum concentration by physical exercise.
- VernacularTitle:ＥＬＩＳＡ法によるＰＧＡＭ‐Ｂの測定と運動による血清濃度の変化
- Author:
TORU TOMIOKA
;
TAKUMITSU HIRO
- Publication Type:Journal Article
- Keywords:
PGAM;
ELISA;
athlete;
exercise;
leaking enzyme
- From:Japanese Journal of Physical Fitness and Sports Medicine
2001;50(3):361-368
- CountryJapan
- Language:Japanese
-
Abstract:
The present paper focuses on the leaking enzyme, PGAM (phosphoglycerate mutase), which is attracting increasing interest among medical researchers, and its relations with physical exercise. As a preliminary, purified PGAM-B was extracted from erythrocyte. First of all, erythrocytes were burst by hypotonic buffer. The contents were let through DEAE-Sepharose column and then through an affinity column bound with PGAM antibody. The purified substance thus obtained is analyzed by SDS-PAGE and has been found only in the single band of approximately 30 kDa. It has been injected to a mouse to develop its antibody. The mouse antibody thus developed and commercial anti r-h-PGAM-B rabbit antibody have been used to in order to perform double antibody sandwich ELISA on PGAM-B. R-h-PGAM-B was used as a standard substance. It has been observed that our standard curve shows a linearity in the range of 0-100ng/100μl, with R2=0.9849. The correla-tion between intensity of physical training and PGAM-B concentration in serum has been analyzed. Non-athlete male students, long distance male and female runners took part in the experiment. The sera of the male runners were found to have 34 times higher PGAM-B concentration than those of the non-athletes. The male runners had 3.6 times higher concentration than those of female runners. Non-athletes ran 10 km and their serum PGAM-B concentration were measured afterwards. It was found that concentration was 3.2 times higher after the run than before it, and this increasing found in all subjects.
