Question

Have a standard curve and determine the concentration of the samples. Get the mean, standard deviation for the two samples tested in triplicate and run a t.test. Is there a difference between the two samples? These were two different sets of serum (two different pigs). I would think they will be different but let's see.

Questions

As you should recall, glucose is a reducing sugar. There are many chemical assays for reducing sugars such as using Nelson's alkaline copper reagent or 3,5 dinitrosalicylic acid (DNS). Why is an enzymatic assay the preferred method for blood glucose?

Convert a typical measurement of 95 mg/dL blood glucose into a mM amount.

The concentration of glucose inside a typical cell is about 1.0 mM. Why is the concentration of glucose inside the cell less than in the blood?

A typical red blood cell has a diameter of about 7 m. Although they are disk-shaped, let's assume they are a sphere and determine the number of glucose molecules in the cell?

Red blood cells are one of the few tissues that rely solely on glycolysis. How much ATP can be made from the amount of glucose in the red blood cell?

Based on questions 2-5, why is it important that the blood glucose concentration remain constant?

Maltose is a disaccharide of glucose (two glucose molecules linked by an alpha 1-4 glycosidic bond). A solution of maltose is tested with the DNS-reagent reducing sugar assay and found to be 50 mM. What would the concentration be measured as if this maltose solution is tested with the glucose-oxidase assay? Explain.

Search the literature, web-sites, wikipedia etc. and find an example of a clinical assay that uses coupled reactions involving an oxidase/peroxidase system. Reference you source and write down the reaction sequence.

Background: Determination of blood glucose is a fundamental test performed in a clinical laboratory. Normal fasting blood glucose levels range from 70-99 mg/dL in healthy adults and hyperglycemia is of course an indication of diabetes mellitus or other medical conditions. One of the more common methods for this determination is to use a glucose oxidase/peroxidase system.
The assay that will be used is called an Endpoint Enzymatic Spectrophotometric assay . Glucose oxidase (GOX) is found in various insects and fungi where it is used as an anti-bacterial agent [4]. GOX oxidizes -D-glucose in into D-gluconolactone with the subsequent production of hydrogen peroxide (eq. 1). Hydrogen peroxide is potent oxidizing agent used by many types of cells to kill pathogens. In the glucose assay the hydrogen peroxide that is released, combined with horseradish peroxidase (HPR), is used to oxidize a dye molecule that is monitored spectrophotometrically (eq. 2).
Many dyes have been used for this assay, but today, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS, figure 1) will be used since it is stable, readily soluble in water and non-toxc

data given
standards 0.726 0.479 0.235 0.139 0.094 0.037 blank
sample 1 0.509 0.48 0.454
sample 2 0.462 0.497 0.539


mg/dL
200
100
50
25
12.5

Answer 1

Question:

Maltose is a disaccharide of glucose (two glucose molecules linked by an alpha 1-4 glycosidic bond). A solution of maltose is tested with the DNS-reagent reducing sugar assay and found to be 50 mM. What would the concentration be measured as if this maltose solution is tested with the glucose-oxidase assay? Explain.

Answer:

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