The normalization of quantitative assay results is critical to the interpretation of proper biological system function. The most common way to evaluate biological system function is through laboratory assays on collected specimens measuring analyte levels correlating to proper system function. Normalization in these assays is essential for generating accurate measurements and assessing proper biological system function.
Normalization in Protein Assays
Assessing total protein content in cell culture or tissue samples utilizing a Bradford assay, (developed by Marion Bradford at the University of Georgia in 1976), allows researchers to characterize cellular health. Protein Assays like, Bradford, Lowry, and other dye-binding assays, have factors such as detergents, chelators, and even protein standards that could cause errors that will adversely affect the result.
We have developed a simple, easy-to-use dual-range BCA Detection Kit (K041-H1) for measuring protein. Our BSA protein standard gives the same result for both BSA and IgG. The kit is highly sensitive measuring proteins at a concentration of 1.68 µg/mL in a variety of sample types for all species. The kit features two 96-well plates and room-temperature stable, liquid reagents that are ready to use.
Normalization in Urine Collection Assays
A urinary collection is a common practice for medical professionals to access proper kidney function. They can do this through a 24-hour urine collection or a single random urine collection. For a 24-hour urine collection, the total volume of urine excreted during a full day is collected and its metabolites are measured. This method is great for accurately measuring total urine output and allows normalization across the collected urine samples. However, the collection and storage of Human urine samples over 24 hours is inconvenient and some analytes may have limited stability in the collected urine volume.
The alternative to a 24-hour collection is to collect a single random urine sample and measure urinary creatinine. Creatinine is a metabolite of phosphocreatine, a molecule used as a store for high-energy phosphate, that can be utilized by tissues for the production of ATP. Creatine and phosphocreatine are converted non-enzymatically to the metabolite creatinine, which diffuses into the blood and is excreted by the kidneys. Under normal conditions, creatinine is created in blood at a constant rate. Its renal excretion is only dependent on urine volume, making creatinine a useful tool for normalizing the levels of other molecules found in urine. In this way, utilizing creatinine to normalize a single random urine sample is far more convenient and accurate than a 24-hour urine collection. Our Urinary Creatinine Kits (K002-H1/H5) allow a rapid, easy-to-use, and precise measurement against a US National Institute of Standards and Technology-calibrated creatinine standard..
Hemoglobin
Normalization in Blood Collection Assays
Blood collection is a common practice for medical professionals to access the proper function of biological systems. Assays on blood specimens can evaluate blood sugar levels, infection, and even hemoglobin function. These assays require normalization to get accurate measurements. Hemoglobin is a reference molecule for the normalization of assays carried out on red blood cells (RBCs), serum and plasma samples. Many enzymes or other molecules assessed in RBCs are reported per gm of hemoglobin. An example would include monitoring enzymes like uridine diphosphate galactose-4-epimerase in patients with galactosemia. Our Dual-Range Hemoglobin Detection Kit (K013-H1) is calibrated to a native human hemoglobin standard and is designed to measure hemoglobin levels in whole blood, serum, plasma, and RBCs. All reagents are stable at 4°C and results are obtained in 20 minutes.