Overview

  • Product name
    Anti-Firefly Luciferase antibody [Luci17]
    See all Firefly Luciferase primary antibodies
  • Description
    Mouse monoclonal [Luci17] to Firefly Luciferase
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-Frmore details
  • Immunogen

    Hybridoma produced by the fusion of spleenocytes from mice immunized with luciferase protein isolated from Photinus pyralis.

  • Positive control
    • WB: Purified luciferase protein. IHC: frozen drosophila embryos
  • General notes

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.08% Sodium Azide
    Constituents: PBS, pH 7.2
  • Concentration information loading...
  • Purity
    Protein A/G purified
  • Clonality
    Monoclonal
  • Clone number
    Luci17
  • Isotype
    IgG1
  • Light chain type
    unknown
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab16466 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
Flow Cyt Use at an assay dependent concentration. PubMed: 18612415

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Target

  • Relevance
    Luciferase from the firefly has become one of the more widely used reporter proteins for the study of gene expression. Luciferase catalyzes a bioluminescent reaction which requires the substrate luciferin as well as Mg2+ and ATP. Mixing these reagents with the cell extract containing luciferase, results in a flash of light that decays rapidly. This light can be detected by a luminometer. The total light emission is proportional to the luciferase activity of the sample.
  • Cellular localization
    Peroxisome
  • Alternative names
    • ec 1.13.12.7 antibody
    • Firefly antibody
    • Luciferase antibody
    • Luciferin 4 monooxygenase antibody

Images

  • All lanes : Anti-Firefly Luciferase antibody [Luci17] (ab16466) at 1 µg/ml

    Lane 1 : Non-transfected 293 whole cell lysate
    Lane 2 : Firefly Luciferase transfected 293 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : NIH3T3 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/10000 dilution

    Performed under reducing conditions.

    Observed band size: 65 kDa
    why is the actual band size different from the predicted?



     

    Lanes 1 - 4: Merged signal (red and green). Green – ab16466 observed at 65 kDa. Red - loading control, ab181602, observed at 37 kDa.

     This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab16466 and ab181602 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • Flow cytometric analysis of HEK293 cells expressing Firefly Luciferase, staining Firefly Luciferase with ab16466.

    Cells were fixed with paraformaldehyde at room temperature for 20 min, rinsed twice with permeabilization buffer (PBS, 1% FBS, 0.1% saponin), and incubated with primary antibody. A FITC-conjugated anti mouse IgG was used as the secondary antibody.

References

This product has been referenced in:
  • Forootan SS  et al. Real-time in vivo imaging reveals localised Nrf2 stress responses associated with direct and metabolism-dependent drug toxicity. Sci Rep 7:16084 (2017). Read more (PubMed: 29167567) »
  • Davis MI  et al. Small Molecule Inhibition of the Ubiquitin-specific Protease USP2 Accelerates cyclin D1 Degradation and Leads to Cell Cycle Arrest in Colorectal Cancer and Mantle Cell Lymphoma Models. J Biol Chem 291:24628-24640 (2016). Read more (PubMed: 27681596) »
See all 12 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Answer

The flow cytometry image shown on ab16466's datasheet is from: Pellegatti P et al. Increased level of extracellular ATP at tumor sites: in vivo imaging with plasma membrane luciferase. PLoS ONE 3:e2599 (2008) as stated in the figure legend.

From the materials section of this paper, it states that cells were incubated with ab16466 (8–10 µg/5×10^5 cells.) I would therefore use this as a starting point for your experiment.

Read More

Answer

Thank you for the clarification. EIther one of these antibodies should work well with your sequence. I personally would select ab16466 because it is a purified antibody rather than an IgG fraction, and therefore will have less chance of non-specific binding. I hope this information is helpful to you. Please let me know if I can be of further assistance.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Schwann Cell)
Loading amount
200 µg
Specification
Schwann Cell
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Oct 16 2012

Answer

Thank you for your enquiry.


Usually the buffer to use for dilutionis the same as your blocking buffer. There are some common buffers that are used which are1-3% BSA buffer, secondary antibody host species serum, Blockase, or even just PBST only.


Please check the protocols on our website for more details.
https://www.abcam.com/index.html?pageconfig=popular_protocols


I hope this information will be helpful. If there is anything else that I can help you with please do not hesitate to contact me.


Have a nice day!

Read More

Answer

Thank you very much for contacting us with your question, and for your patience while I have been in touch with the lab regarding these antibodies.

Unfortunately, the sensitivities of these two antibodies has not been determined at this time.

I am sorry that I do not have further information for you, but if there is anything else that we can do for you please let me know and I'll be happy to help.

Read More

Answer

Thank you for contacting Abcam

We have a western blot image, which is attached showing the antibody with recombinant luciferase. We have not done further testing to see what is either smallest concentration of antibody that is needed to recognize a know concentration of the protein.

If there is anything else I can help you with, please let me know.

Read More

Answer

I hope youmanaged to open the attachment file I sent previously. The image contains the list of non-conjugated HRPanti-Firefly antibodies we have in our catalogue, including goat polyclonal, chicken polyclonal, and rabbit polyclonal antibodies. These antibodies can be used in WB application, so I am sure you will be able to find one suitable for your experiment.



Our current stock status for these antibodies are as below:



ab498 Goat polyclonal to Firefly Luciferase (1 local stock)

ab81823 Goat polyclonal to Firefly Luciferase (1 local stock)

ab99944 Rabbit polyclonal to Firefly Luciferase (8 overseas stock, lead time 1 week)

ab18595 Chicken polyclonal to firefly Luciferase (no stock, lead time 1-2 weeks from order)



I hope the stock information will help you decide on which product to use for your experiment.



Good luck with your researchand have a nice day!

Read More

Answer

Thank you for your enquiry.



We do have a few Firefly antibodies in our catalogue (see attached file)but unfortunately, none of them have been tested in IP application. These products however can be used in WBand might also work in IP.For your information, one of our best selling antibodies is ab16466 Mouse monoclonal [Luci17] to Firefly Luciferase.



Please rest assured that all tested applications are specified on Abcam product datasheets, and these are updated as soon as any new information is brought to our attention. Although IP is not stated on the datasheet, this does not mean that the antibody will not work, just that we can not guarantee that you will get good results. Please rest assured that if we have tested it in IP and found that it does not work, the application will be stated on the datasheet as not suitable for the product.



Regarding ourrecommended IP protocol, please see the following link for more information.



https://www.abcam.com/index.html?pageconfig=resource&rid=11385



I hope this information will still be helpful.If there is anything else that I can help you with please do not hesitate to contact me.



Have a nice day!

Read More

Answer

Thank you for your enquiry and your interest in our products.

Luciferase from the firefly and the luciferase genes can be synthesized and inserted into organisms or transfected into cells. Normally, monkeys do not express luciferase unless the samples contain transfected cells.

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Answer

Thank you for contacting Abcam.

This product was created using the whole purified protein and an immunogen. The fusion of spleenocytes is an antibody production technique for creating monoclonals.

The epitope for this antibody has not yet been mapped.

To our knowledge this product has not been tested for inhibition of enzymatic activity.

I hope this information is helpful. Please let me know if you have any questions.

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1-10 of 13 Abreviews or Q&A

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

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