Anti-Firefly Luciferase antibody [3i1] (ab64564)

Overview

  • Product nameAnti-Firefly Luciferase antibody [3i1]
    See all Firefly Luciferase primary antibodies
  • Description
    Mouse monoclonal [3i1] to Firefly Luciferase
  • Tested applicationsSuitable for: WB, ELISA, ICCmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Firefly
  • Immunogen

    Full-length recombinant luciferase from Photinus pyralis.

  • Positive control
    • 3T3 cell lysate containing 5ng of recombinant luciferase

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Add glycerol to a final volume of 40% for extra stability and aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage bufferPreservative: 0.05% Sodium Azide
    Constituents: 0.1M Tris glycine, 150mM Sodium chloride, pH 7.4
  • Concentration information loading...
  • PurityProtein G purified
  • ClonalityMonoclonal
  • Clone number3i1
  • IsotypeIgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab64564 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
ELISA
ICC
  • Application notesELISA: Use at an assay dependent dilution.
    ICC: Use at an assay dependent dilution.
    Is unsuitable for IHC-P.
    WB: Use at a concentration of 0.2 - 5 µg/ml. Predicted molecular weight: 66 kDa.
    Note: 0.5-2 µg/ml detects 5ng of recombinant luciferase that was added to 3T3 cell lysate.

    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • RelevanceLuciferase 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 localizationPeroxisome
    • Database links
    • Alternative names
      • ec 1.13.12.7 antibody
      • Firefly antibody
      • Luciferase antibody
      • Luciferin 4 monooxygenase antibody

    Anti-Firefly Luciferase antibody [3i1] images

    • All lanes : Anti-Firefly Luciferase antibody [3i1] (ab64564) at 5 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
      Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size : 66 kDa
      Observed band size : 65 kDa (why is the actual band size different from the predicted?)

      Lanes 1 - 4: Merged signal (red and green). Green – ab64564 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 ab64564 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.

    References for Anti-Firefly Luciferase antibody [3i1] (ab64564)

    ab64564 has not yet been referenced specifically in any publications.

    Product Wall

    Application Western blot
    Loading amount 0.05 µg
    Gel Running Conditions Reduced Denaturing (4-15% Tris-glycine)
    Sample Firefly (Photinus pyralis) Recombinant protein (QuantiLum Recombinant Luciferase (Promega))
    Specification QuantiLum Recombinant Luciferase (Promega)
    Treatment Heated in the presence of sHSP followed by UV crosslinking
    Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Username

    Miss. Rachel Loh

    Verified customer

    Submitted Jan 13 2015

    We have no firefly luciferase antibody tested in IP which is mouse or rabbit origin. Only ab498- Goat polyclonal to firefly Luciferase Other Firefly Luciferase antibodies tested in WB: 1. ab16466 Firefly Luciferase antibody [Luci17] Mouse monocl...

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