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Nitric Oxide Assay Kit (Fluorometric) (ab65327)

Q&A (17)Specific References (1)

Overview

  • Product nameNitric Oxide Assay Kit (Fluorometric)See all Nitric oxide kits ...
  • Tests
    200 x 1 assay
  • Sample type
    Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay typeQuantitative
  • Assay time
    3h 00m
  • Product overview

    Abcam's Nitric Oxide Assay Kit (Fluorometric) provides an accurate, convenient measure of total nitrate/nitrite concentration in a simple two-step process. In the first step nitrate is converted to nitrite by nitrate reductase. In the second step, nitrite reacts with the fluorescent probe DAN (2, 3 diaminonaphthalene). NaOH enhances the fluorescent yield. The fluorescent intensity is proportional to the total nitric oxide production. The kit has been tested with culture media, plasma, and tissue homogenates.
    Visit our FAQs page for tips and troubleshooting.

     

    If you are interested in a colorimetric detection kit, please check Nitric Oxide Assay Kit (Colorimetric) (ab65328).

  • Notes

    Nitric oxide (NO) plays an important role in neurotransmission, vascular regulation, immune response and apoptosis. Since NO is rapidly converted to nitrite (NO2-) and nitrate (NO3-), the total concentration of nitrite and nitrate is used as a quantitative measure of NO production.

  • Tested applicationsFunctional Studies more details

Properties

  • RelevanceNitric 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 ab65327 in the following tested applications.

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

    Application Notes
    Functional Studies FuncS

    Nitric Oxide Assay Kit (Fluorometric) images

    • Functional Studies - Nitric Oxide Fluorometric Assay Kit (ab65327)
      Functional Studies - Nitric Oxide Fluorometric Assay Kit (ab65327)
      Nitrite, nitrate assay in the presence and absence of nitrate reductase. Assays were performed according to the kit protocol with 1 hour conversion of nitrate to nitrite at Step 5.

    Protocols

    References for Nitric Oxide Assay Kit (Fluorometric) (ab65327)

    This product has been referenced in:
    • Huang Z  et al. Stimulation of Unprimed Macrophages with Immune Complexes Triggers a Low Output of Nitric Oxide by Calcium-dependent Neuronal Nitric-oxide Synthase. J Biol Chem 287:4492-502 (2012). Read more (PubMed: 22205701) »

    See 1 Publication for this product

    Publishing research using ab65327 ? Please let us know so that we can cite the reference in this datasheet

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    Displaying 1 - 10 of 17 results for Abreviews and Q&A

    10
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    how should cell lysate be prepared?
    can 1% TX-100 -based lysis buffer be used? Is it compatible with kit?


    The lab has confirmd that your lysis buffer would be compatible, and that pretty much any generic lysis buffer with a mild detergent will work.

    we had your Nitric Oxide Assay Kit (Fluorometric) (ab65327). However when we look at kit protocol, i couldnt see any tissue preparation method. we will use this kit for measurement of NO in retina and we need tissue praparation method. because in florimet...

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    Thank you for contacting us.

    For tissue samples: Start with 10-50 mg of the tissue, add 500-1000 μl (or ˜3-4 volumes of tissue size) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lys...

    Read More

    Hi,
    I would like some suggestions on which product would suit best for my work. I want to trace the amount of nitric oxide in freeze specimens and paraffin specimens. I've found one of your assay kit 'ab65327Nitric Oxide Assay Kit (Fluorometric)', bu...

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    Thank you for your enquiry regarding ab65327.

    We have nottested this kit in frozen tissue in our lab. However, if the samples were frozen appropriately, there is still a possibility that this kit might work on lysates from such tissues.
    Read More

    I don't quite understand what you mean by using fresh medium. It is fresh medium that I used for standard curve, and it's just plain medium, not from tissue culture which I would use for my test. Deproteinization is suggested by the prot...

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    Thank you for your reply.
    I am sorry that you are not happy with this kit. Please see below for the refund information.

    The deproteinization step would apply for the samples, not for the standard curve.

    The lab also send me the f...

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    It is suggested by the protocol that if the culture medium contains phenol red, it's ideal to add it into the standard curve as phenol red will affect the reading. So in the 0pmol/well, it also contains same amount of diluted medium.
    It&...

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    Thank you for your email.

    As I had mentioned in a previous email, I would be happy to replace the kit for you. Can you please confirm thatthis is what you prefer or rather a credit or refund?

    On the same hand, it is important to us to ...

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    How long will it take? I thought I'm able to get a replacement.

    At the moment, we do not know if there is something wrong with the kit (as the lab tested the assay buffer and it was fine) or with the way the samples were prepared or measured.

    Can you please let me know what exactly was added to the wells wit...

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    Group A-C is done at different times with serial dilution of the mediums. They're standard curve, and I didn't do standard curve in triplicate. Colume 1-6 refers to different concentration of the nitrate standards and they are: 0, 200,40...

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    Thank you for your email.
    I will pass the data and information to the lab for evaluation.

    In the meantime, they have checked the assay buffer andit did not produce any RFU in their QC testing done this morning. Thus, the buffer seems to be ...

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    sorry for the delay. I just got the information from the purchasing office, here's the information:
    PO#: xxx
    Lot#: GR94531-1
    Please also find the attachment for the result.

    Thank you for the information. I found your order in the system.


    Could you please clarify what Group A, B, and C stand for or maybe how they differ? Are these the same samples in triplicate?
    Column 1 thru 6 contain data, but what is t...

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    used cell culture medium (EBM2, EGM2)
    high background in standrad curve
    blank (assay buffer) readout = 14,000 RFU
    all other values are higher and become saturated

    Thank you for contacting us.

    I heard back from the lab and they will check the assay buffer, but they would need to know the lot number you have. If you please could send me your order or PO number (or lot number on the kit), I will let them kn...

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    Inquiry: Dear Technical Support Team: I would like to know if Nitric Oxide Assay Kit (Fluorometric) (ab65327) is compatible with liver tissues that have been stored at -80C˚ after snap frozen. If it is compatible with frozen liver tissues, do you suggest ...

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    Yes, liver frozen in the manner you desribe can be used for this assay. We don’t have a specific recommended homogenization buffer for tissue.


    Hope this information has been helpful for you. Please let me know if you have any other q...

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