Overview

  • Product name
    Nitric Oxide Assay Kit (Colorimetric)
    See all Nitric oxide kits
  • Sample type
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate
  • Assay type
    Quantitative
  • Sensitivity
    1 nmol/well
  • Range
    1 nmol/well - 10 nmol/well
  • Assay time
    1h 30m
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mammal
  • Product overview

    Abcam's Nitric Oxide Assay Kit (Colorimetric) provides an accurate, convenient measure of total nitrate/nitrite in a simple two-step process. The first step converts nitrate to nitrite utilizing nitrate reductase. The second step uses Griess Reagents to convert nitrite to a deep purple azo compound. The amount of the azochromophore accurately reflects nitric oxide amount in samples.
    Visit our FAQs page for tips and troubleshooting.

    If you are interested in a fluorometric detection kit, please check Nitric Oxide Assay Kit (Fluorometric) (ab65327).

  • Notes

    Nitric oxide (NO) plays an important role in neurotransmission, vascular regulation, immune response and apoptosis. NO is rapidly oxidized to nitrite and nitrate which are used to quantitate NO production.

  • Tested applications
    Suitable for: Functional Studiesmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components Identifier 2 x 96 tests 200 tests
    Assay Buffer WM 1 x 30ml 1 x 30ml
    Enhancer (Lyophilized) Purple 1 vial 1 vial
    Enzyme Cofactor (Lyophilized) Blue 1 vial 1 vial
    Griess Reagent R1 NM 1 x 10ml 1 x 10ml
    Griess Reagent R2 Amber NM 1 x 10ml 1 x 10ml
    Microtiter plate 2 units 2 units
    Nitrate Reductase (Lyophilized) Green 1 vial 1 vial
    Nitrate Standard (Lyophilized) Yellow 1 vial 1 vial
    Nitrite Standard (Lyophilized) Orange 1 vial 1 vial
    Plate Cover 2 units 2 units
  • Research areas
  • Relevance
    Nitric oxide (NO) is a key vertebrate biological messenger, playing an important role in neurotransmission, vascular regulation, immune responses and apoptosis. NO , also known as "endothelium-derived relaxing factor" or "EDRF", is synthesized from L-arginine, oxygen and NADPH by various NO synthases. Most of the NO in the cell is oxidized to nitrite (NO2-) and nitrate (NO3-), and therefore the concentrations of these anions are generally as a quantitative measure of NO production.

Applications

Our Abpromise guarantee covers the use of ab65328 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent dilution.

Images

  • Nitrite and nitrate measured in cell lysates showing quantity (micromol) per 106 cells

  • Nitrite and nitrate measured in biologicals showing concentration (micromolar)

  • Standard curve (colorimetric) : mean of duplicates (+/-SD) with background readings subtracted

  • Standard curve (Colorimetric) : mean of duplicates (+/-SD) with background readings subtracted

Protocols

References

This product has been referenced in:
  • Olivares Pacheco J  et al. Metabolic Compensation of Fitness Costs Is a General Outcome for Antibiotic-Resistant Pseudomonas aeruginosa Mutants Overexpressing Efflux Pumps. MBio 8:N/A (2017). Functional Studies ; Pseudomonas aeruginosa . Read more (PubMed: 28743808) »
  • Gammoh O  et al. Chlorpheniramine and escitalopram: Similar antidepressant and nitric oxide lowering roles in a mouse model of anxiety. Biomed Rep 6:675-680 (2017). Read more (PubMed: 28584640) »

See all 10 Publications for this product

Customer reviews and Q&As

Merci pour votre patience.

Le laboratoire vient de me confirmer que le NO est stable et que les échantillons de surnagéant aussi peuvent donc être conservées en aliquots à-20°C ou -80°C.

J'e...

Read More

Thank you for contacting us. The Fluorometric assay kit ab65327 is much more sensitive (about 10x) than kit ab65328 (colorimetric assay kit). For ab65328 (colorimetric): The absorbance of samples should be in the linear range of the stan...

Read More

Thank you for your enquiry. This kit will be compatible with MEM with phenol red. Media that contain phenol red as a pH indicator do not interfere with the Griess reaction as the indicator is typically yellow colored under the conditions of the Gri...

Read More

fter adding Griess Reagent R2 in standard and unknown leave it for 10 minutes at room temperature. The color developed will be stable for an hour.

Abreviews
I used this kit to measure nitric oxide (nitrate and nitrite combined) in the plasma of adult male red-winged blackbirds (Agelaius phoeniceus) housed in an outdoor aviary. Preliminary data suggests that the assay requires a minimum of 15uL of plasma per well (ie, 30uL plasma to run in duplicate) to produce repeatable results. Thus far, we have found birds to have levels of NOx between 14-250 uM. It appears that birds with an active infection could have elevated NOx.
Preliminary data also suggests that error can be easily introduced when ready-ing the samples for the assay by deproteinating plasma with PCA. I suspect that this is particularly problematic for samples with absorbance reading less than 0.2, but we need more evidence to be certain.
I did notice that the instructions on the web site were not the same as in the booklet sent. In the booklet I received, the instructions said to use the plate reader after adding the second Griess reagent with no wait time listed. On the online book an incubation time was listed (10 min). There were also inconsistencies in the the list of reagents provided and what was actually provided (We got plates, which is great!). The plates were labeled with a store in -20C, even though they're just plastic plates that isn't necessary.
Technical support was very helpful in answering questions quickly whenever I called - was really happy with the support.
Username

Abcam user community

Verified customer

Submitted Feb 01 2016

Yes, heparin should be fine to use with ab65328 and ab118970.



Standard amounts of salts, Triton and protease inhibitors should be alright. We have not tested this kit’s function using high salt/detergent but theoretically small amounts of these should not interfere with the assay.



I am happy to confirm that extra assay buffer can now be purchased using the following product number:

ab171368

http://www.abcam.com/index.html?datasheet=171368 (or use the following: http://www.abcam.com/index.html?datash...

Read More


This kit does not include a step to remove proteins. Depending on your sample type, it is however recommended to use 10kD spin columns (ab93349) to deproteinate your samples.

For example, it is recommended for serum samples since there a...

Read More

Thank you for contacting us.


We have not tried boiling the end products to see if it affects the readings, but I would guess that it would. I would think that the safest way to use these samples and maintain the analysis efficiency would ...

Read More

1-10 of 28 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up