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Anti-Firefly Luciferase 抗体 [Luci17] (ab16466)

Reviews (5)Q&A (10)References (22)
Western blot - Anti-Firefly Luciferase antibody [Luci17] (ab16466)

    Key features and details

    • Mouse monoclonal [Luci17] to Firefly Luciferase
    • Suitable for: WB
    • Reacts with: Firefly
    • Isotype: IgG1

    製品の概要

    • 製品名

      Anti-Firefly Luciferase antibody [Luci17]
      Firefly Luciferase 一次抗体 製品一覧

    • 製品の詳細

      Mouse monoclonal [Luci17] to Firefly Luciferase

    • 由来種

      Mouse

    • アプリケーション

      適用あり: WBmore details

    • 種交差性

      交差種: Firefly

    • 免疫原

      Recombinant fragment corresponding to Firefly Firefly Luciferase. Hybridoma produced by the fusion of splenocytes from mice immunized with luciferase protein isolated from Photinus pyralis.
      Database link: P08659

    • ポジティブ･コントロール

      • WB: Purified luciferase protein

    • 特記事項

       Analysis of gene expression is most commonly assayed by transient transfection. Systems are generally based on the use of fusion genes which are inserted into cells, and the gene expression is assayed within 48 hours after introduction of DNA. Usually the fusion consists of the promoter binding site or enhancer sequence under study which is attached to a reporter gene. The amount of the reporter protein synthesized under the experimental conditions, is presumed to reflect the ability of the sequences studied to direct or promote transcription. Several enzymes are commonly used as reporter proteins, among them are chloramphenicol acetyl transferase (CAT), -galactosidase, human growth hormone (hGH) and luciferase. Luciferase has become one of the widely used reporter enzymes. The enzyme catalyzes a bioluminescent reaction which requires the substrate luciferin as well as Mg+2 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. The use of an antibody to detect luciferase can provide an alternative detection assay which directly detects luciferase protein levels, and thus has the advantage that it does not require luciferase activity and is not dependent on rapid kinetics. Moreover, antibodies can detect the luciferase enzyme expression in situ, providing a means to study the localized signal sequences using luciferase as a reporter gene. Reacts with Luciferase (Firefly) Protein.

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    製品の特性

    • 製品の状態

      Liquid

    • 保存方法

      Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

    • バッファー

      pH: 7.20
      Preservative: 0.08% Sodium azide
      Constituent: PBS
    • Concentration information loading...

    • 精製度

      Protein A/G purified

    • ポリ/モノ

      モノクローナル

    • クローン名

      Luci17

    • アイソタイプ

      IgG1

    • 軽鎖の種類

      unknown

    • 研究分野

    アプリケーション

    The Abpromise guarantee

    Abpromise保証は、 次のテスト済みアプリケーションにおけるab16466の使用に適用されます

    アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

    アプリケーション Abreviews 特記事項
    WB (2)
    Use at an assay dependent concentration.
    特記事項
    WB
    Use at an assay dependent concentration.

    ターゲット情報

    • 関連性

      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.

    • 細胞内局在

      Peroxisome

    • 参照データベース

      • 別名

        • ec 1.13.12.7 antibody
        • Firefly antibody
        • Luciferase antibody
        • Luciferin 4 monooxygenase antibody

      画像

      • Western blot - Anti-Firefly Luciferase antibody [Luci17] (ab16466)
        Western blot - Anti-Firefly Luciferase antibody [Luci17] (ab16466)
        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.

      参考文献 (22)

      ab16466 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

      ab16466 は 22 報の論文で使用されています。

      • Pang HH  et al. Bioengineered Bacteriophage-Like Nanoparticles as RNAi Therapeutics to Enhance Radiotherapy against Glioblastomas. ACS Nano 17:10407-10422 (2023). PubMed: 37120837
      • Ling Z  et al. Targeting CCL2-CCR4 axis suppress cell migration of head and neck squamous cell carcinoma. Cell Death Dis 13:158 (2022). PubMed: 35177591
      • Deplazes S  et al. Intrabiliary infusion of naked DNA vectors targets periportal hepatocytes in mice. Mol Ther Methods Clin Dev 27:352-367 (2022). PubMed: 36381301
      • Cheng K  et al. Bioengineered bacteria-derived outer membrane vesicles as a versatile antigen display platform for tumor vaccination via Plug-and-Display technology. Nat Commun 12:2041 (2021). PubMed: 33824314
      • Vichas A  et al. Integrative oncogene-dependency mapping identifies RIT1 vulnerabilities and synergies in lung cancer. Nat Commun 12:4789 (2021). PubMed: 34373451
      View all Publications for this product

      レビューと Q&A

      Show All レビュー Q&A
      レビューを送る 質問を送る

      1-10 of 15 Abreviews or Q&A

      Immunohistochemistry (Frozen sections) abreview for Anti-Firefly Luciferase antibody [Luci17]

       Inconclusive
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Frozen sections)
      Sample
      Mouse Tissue sections (Brain)
      Permeabilization
      Yes - 0.1% Triton-X
      Specification
      Brain
      Blocking step
      BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 21°C
      Fixative
      Formaldehyde
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Brenda Lam

      Verified customer

      投稿 Oct 03 2022

      Immunohistochemistry (Frozen sections) abreview for Anti-Firefly Luciferase antibody [Luci17]

       Below Average
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Frozen sections)
      Sample
      Mouse Tissue sections (Brain tumor (DIPG))
      Permeabilization
      Yes - 0.5% triton
      Specification
      Brain tumor (DIPG)
      Blocking step
      Goat serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 4°C
      Fixative
      Paraformaldehyde
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      投稿 Dec 03 2020

      Western blot abreview for Anti-Firefly Luciferase antibody [Luci17]

       Excellent
      Abreviews
      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
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      投稿 Oct 16 2012

      Question

      Primary antibody dilutions are not specified either in the website or in the paper that used this antibody for flow cytometry and are cited in the website.

      Could you give me this information so I can have a starting point?

      Read More
      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

      Question

      We are using pGL3 Luciferase Reporter Vector. It has a modified coding region for firefly (Photinus pyralis) luciferase that has been optimized for monitoring transcriptional activity in transfected eukaryotic cells.


      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

      Question

      I have purchased an antibody (ab16466) which is 1mg/ml and i have to make 400µg/ml, can you tell me what buffer do i need to dilute it.

      Read More
      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

      Question

      Good Afternoon abcam Technical Service,
      I am looking to find out what the sensitivity of your two monoclonal antibodies: Anti-firefly Luciferase antibody [Luci 21 1-107] (ab7358) and Anti-Firefly Luciferase antibody [Luci17] (ab16466) is?

      Thank you in advance for your help.

      Sincerely,

      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

      Question

      What is the sensitivity of this antibody in a western blot against a recombinant protein in western blot?

      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

      Question

      Thanks for your quick reply. Now i know that you have ab16466 Mouse monoclonal [Luci17] . I just wanted to know if you have any-other anti-firefly antibody raised not in mouse.

      So basically i want to to do is pull-downs with Mouse monoclonal [Luci17] and do a western with anti-firefly raised either in goat or rabbit (if possible with HRP for ECL but not required).

      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

      Question

      I need to find out if you have anti-firefly antibody for IP and do a western subsequently. If yes can you send me a protocol for IP with anti-firefly antibody.

      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

      1-10 of 15 Abreviews or Q&A

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