Medición de la concentración de a2-macroglobulina (a2m) en el suero de la rata.

ALFREDO RIGALLI

Rosario

Congreso; VI Congreso y XXIV Reunión Científica anual de la Sociedad de Biología de Rosario.; 2004

Sociedad de biología de Rosario

In rats, a2M is an acute phase reaction glycoprotein. It inhibits proteinases and binds cytokines and growth factors. Sodium monofluorophosphate (MFP), a drug used in the treatment of osteoporosis and the prevention of caries, binds to a2M, forms a complex and modifies the antiproteasic activity. There are several techniques for measuring a2M, but they permit to obtain relative values, and they are time-consuming or technologically inapplicable. The aim of this work was to develop a method for the measurement of plasmatic levels of a2M in the rat. The proposed technique is based on the dot-blot technology for protein measurement. Basically, it consists of: 1- The binding of serum proteins to a nitrocellulose membrane and subsequent blocking of nonespecific sites 2- The incubation with primary antibody to rat a2M. 3- The incubation with antiGuinea Pig Immunoglobulin G alkaline phosphatase conjugated secondary antibody. 4- The development of dots with a specific compound that produces a blue cromophore. 5- The digitalization of the blue area. 6- The measurement of intensity (I) and area (A) of the dot with a specific software. The technique describe above was applied to solutions with known concentrations of rat a2M. A and I were measured and AxI was calculated. The linear regression between AxI and a2M concentrations was statistically significant (p<0,0001). The coefficient of variation for a serum pool of normal rats was 11.5%. Conclusions: 1- This technique allows to measure rat a2M concentration. 2- Results can be obtained in approximately 6 hours. 3- The complexity and the cost of the method are low.