BioMedical Nitrate & Nitric Oxide Assays - with Superior Stock Nitrate Reductase Inside!

Bio-Medical Nitrate Assays
Nitrate-Reductase Based!

Superior Stock Nitrate Reductase Inside!

    The Nitrate Elimination Co., Inc.
    Green Chemistry for Nitrate Detection
                             Biotechnology People Can Use

Revised and Updated: 12/17/2008 1.888.NITRATE www.nitrate.com

Nitric Oxide (NO) Produced in Body Tissues and Fluids Results
in Residual Nitrate in the Human Body

NO produced in response to infection and stress raises the Nitrate level over that normally found in the serum. This is due to conversion of NO to nitrate and nitrite. Low levels of nitrate in serum are normal due to hormonal NO production and dietary intake. However, when the body is being challenged, the NO is often produced and results in an elevated level of nitrate, which can be viewed as an indicator of increased production of NO.

In research studies following NO levels, the transiently produced NO gas is difficult to detect unless expensive instrumentation is available. Since NO is stoichiometrically converted to nitrate and nitrite, the levels of the NO produced in a response can be tracked by assaying for nitrate and nitrite. The easiest test for nitrate is done by converting it to nitrite by enzymatic reduction catalyzed by nitrate reductase. The nitrite is detected by a simple colorimetric assay using the Griess Reagents. This approach works well in tissue culture based studies and isolated body tissues and fluids.

NECi Lab Nitrate Test Kits (Lab-NTK) use small volumes of sample (5 to 500 µL) and generally have no problem with interferences due to protein content of the samples. Micro Plate Format is available for investigators with Micro Plate Readers. Test Tube Format is available for investigators with a colorimeter or spectrophotometer. NADH: Nitrate Reductase is used in NECi Lab-NTK, the reagents are less costly and the kits cheaper. Lab-NTK are highly sensitive, reproducible and accurate. To improve linearity of the Nitrate Standard Curve, a quench agent is provided to remove excess NADH from low nitrate sample.

BioMedical Literature on Enzyme-Based Nitrate Assays for NO Research:

Granger DL, RR Taintor, KS Broockvar & JB Hibbs (1996) Measurement of nitrate and nitrite in biological samples using nitrate reductase and Griess reaction. Methods in Enzymology 268: 142-151
Grisham, MB, GG Johnson, & JR Lancaster (1996) Quantitation of nitrate and nitrite in extracellular fluids. Meth. Enzymol. 268: 237-245
Guevara, I, J Iwanejko, A Szczudlik (1998) Determination of nitrite/nitrate in human biological material by the simple Griess reaction. Clin Chim Acta 274:177-88.
Rao, AM, A Dogan, JF Hatcher, RJ Dempsey (1998) Fluorometric assay of nitrite and nitrate in brain tissue after traumatic brain injury and cerebral ischemia. Brain Res 793:265-270. Some features of this method are incorporated in NECi Micro Plate Nitrate Test Kits.

NECi Lab Nitrate Test Kits

Based on the Enzyme - Superior Stock Nitrate Reductase

Small Sample Volume: 5 to 500 µL

2 Formats: Micro Plate and Test Tube

2 Nitrate Ranges: Low = 3.6 to 71.4 µM & Std = 36 to 714 µM

Potential Applications:

Nitric Oxide (NO) Research

Organ Transplant Studies

Sepsis Indicator

Low Grade Infections

Inflammation Indicator

No Sample Preparation for:

Tissue Culture Media

Blood Plasma

Serum

Urine

NECi Lab and Micro Plate Format Nitrate Test Kits (NTK)
Call 1-888-NITRATE (648-7283) to Order on-Line

NECi Lab Nitrate Test Kits (L-NTK)
Less expensive than most other commercial NO/Nitrate Test Kits

Micro Plate Nitrate Test Kits (M-NTK)
96 Well Micro Plate Format
You need a Micro Plate Reader!

Low Range (3.6 to 71.4 µM Nitrate):
Catalog No. M-NTK-302
Sample Size = 50 µl or 0.05 ml -- 96 Well Micro Plate provided
Standard Range (35.7 to 714 µM Nitrate):
Catalog No. M-NTK-301
Sample Size = 5 µl or 0.005 ml -- 96 Well Micro Plate provided

Test Tube Format -- Colorimeter Required
The Only Test Tube Nitrate Test Kit Available for
Bio-Medical Research

Standard Range (35.7 to 714 µM Nitrate/50 µL Sample):
A. Catalog No. L-NTK-201
25 Determinations using a Colorimeter in your test tubes
B. Catalog No. L-NTK-203
100 Determinations using a Colorimeter in your test tubes
Low Range (3.6 to 71.4 µM Nitrate/500 µL Sample):
A. Catalog No. L-NTK-202
25 Determinations using a Colorimeter in your test tubes
B. Catalog No. L-NTK-204
100 Samples using a Colorimeter using your test tubes