Overview

  • Product name
  • Description
    Rabbit polyclonal to NOX3
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human NOX3.

    Read Abcam's proprietary immunogen policy (Peptide available as ab101218.)

  • Positive control
    • This antibody gave a positive signal in HeLa and HEK293 whole cell lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab81864 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 62 kDa (predicted molecular weight: 64 kDa).
ICC/IF Use a concentration of 1 µg/ml.

Target

Images

  • All lanes : Anti-NOX3 antibody (ab81864) at 1 µg/ml

    Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 64 kDa
    Observed band size: 62 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 105 kDa, 85 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 20 minutes


    Abcam recommends that this antibody is blocked with milk rather than BSA.
  • ICC/IF image of ab81864 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab81864, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% MCF7 fixed (10 min) MCF7 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa cells at 1µg/ml

References

This product has been referenced in:
  • Plantinga TS  et al. Rare NOX3 Variants Confer Susceptibility to Agranulocytosis During Thyrostatic Treatment of Graves' Disease. Clin Pharmacol Ther N/A:N/A (2017). Human . Read more (PubMed: 28486791) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you for contacting us this morning. As we discussed all the antibodies mentioned have been used with human samples and have been tested in Western blotting and would therefore be covered by the Abpromise if used under these conditions. More information on the Abpromise can be found here: www.abcam.com/abprimise If you find that the antibodies are not performing as expected then we would have a look at how you have been using the antibody. This is to find out if there is any advice we can give in order to help, as well as for quality control purposes so we can find out why the antibody is not working as expected and any additional testing we may need to implement. If the antibody is not performing as expected then we can provide a replacement antibody, a refund or a credit note. I have had a look into the positive controls which may be used with the antibodies. Currently for the antibodies we recommend as follows: ab61248: Human kidney lysate ab80508: MCF7 cell lysate ab81864: HeLa and HEK293 whole cell lysates ab110400: Extracts from Jurkat cells Unfortunately from reviewing the literature I could not find any tissue that could guarantee the expression of all four proteins, especially NOX3 and 5: http://www.nih.go.jp/JJID/57/S28.pdf http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2763943/pdf/nihms-134001.pdf However, if you are confident that the proteins are expressed in the tissue of interest (in lab testing or from publication) or you use a positive control of some sort then there will be no problem in obtaining an alternative antibody or refund if the antibodies are found to not be performing as expected. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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