Determination of serum protein concentration in nanoliter blood samples using fluorescamine or o-phthalaldehyde
✍ Scribed by John W. Viets; William M. Deen; Julia L. Troy; Barry M. Brenner
- Publisher
- Elsevier Science
- Year
- 1978
- Tongue
- English
- Weight
- 572 KB
- Volume
- 88
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
Fluorometric procedures were developed to permit measurement of total protein concentration in nanoliter serum samples, using either fluorescamine or ophthalaldehyde. The sensitivities of assays using these two reagents were similar, but the o-phthalaldehyde method was found to be somewhat simpler and more reproducible. Accurate measurements could be obtained on serum samples of 4 to 5 nl with either reagent, by using a serum standard. Fluorescence differed considerably among individual serum proteins, albumin generally showing greater fluorescence than globulins. Small molecular weight species in serum did not contribute appreciably to total serum fluorescence with either reagent.
Methods
Fluorescamine was obtained from Roche Diagnostics (Nutley, New Jersey); o-phthalaldehyde, Brij (polyoxyethylene lauryl ether), and 2-mercaptoethanol were from Pierce Chemical Co. (Rockford, Illinois); bovine serum albumin, bovine y-globulins (Cohn Fraction II), and bovine mixed globulins (Cohn Fraction III) were from Sigma Chemical Co. (St. Louis,