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Core Laboratory

17. Human Adiponectin (Linco Res. Inc.)

Adipocyte complement-related protein of 30 kDa (ACRP30) is a secreted serum protein expressed exclusively in differentiated adipocytes. Recent studies have indicated that its expression and serum levels are reduced in humans and animals with obesity and insulin resistance. Metabolic studies have demonstrated a role for ACRP30 in the regulation of glucose and lipid homeostasis.

In this RIA kit, there is competition between labeled tracer and unlabeled antigen for the limited and constant number of binding sites on the antibody. Thus, the amount of tracer bound to antibody will decrease as the concentration of unlabeled antigen increases. This can be measured after separating antibody-bound from free tracer and counting one or the other, or both fractions. Adiponectin RIA assay utilizes ¹²⁵I-labeled Murine Adiponectin and a Multispecies Adiponectin Rabbit antiserum to determine the level of Adiponectin in serum, plasma or tissue culture media by the double antibody/PEG technique.

Acceptance Criteria:

1. The run will be considered accepted when all Quality Control values fall within the calculated Quality Control Range; if any QC's fall outside the control range, review results with supervisor.
2. If the difference between duplicate results of a sample is >10% CV, repeat the sample.
3. The limit of sensitivity for the Human Adiponectin assay is 1 ng/ml (100 µl sample size).
4. The limit of linearity for the Human Adiponectin assay is 200 ng/ml (100 µl sample size). Any result greater than 200 ng/ml should be repeated on dilution using Assay Buffer as a diluent.

References:

1. Thorell, J.I. Scand. J. Clin. Lab. Invest. 31:187, 1973.
2. Feldman, H. and Rodbard, D. “Mathematical Theory of Radioimmunoassay”, in:W.D Odell and Doughaday, W.H. (Ed.), Principles of Competitive Protein-Binding Assays. Philadelphia: J.B. Leppincott Company; pp 158-203, 1971.
3. Westgard, J.O., et. al. A multi-rule Shewhart chart for quality control in clinical chemistry. Clin. Chem. 27:493-501, 1981.

 

For more information, please contact the Core Laboratory manager or director.