Recombinant
RabMAb

Recombinant Anti-Firefly Luciferase antibody [EPR17790] (ab185924)

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Overview

  • Product name

    Anti-Firefly Luciferase antibody [EPR17790]
    See all Firefly Luciferase primary antibodies

  • Description

    Rabbit monoclonal [EPR17790] to Firefly Luciferase

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Firefly
    Does not react with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Firefly Firefly Luciferase aa 150-250. The exact sequence is proprietary.
    Database link: P08659

  • Positive control

    • WB: Firefly Luciferase transfected HEK293 whole cell lysate. ICC/IF: Firefly Luciferase transfected HEK293T cells.

  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR17790

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab185924 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.
ICC/IF 1/500.
WB 1/20000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).

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

  • Database links

    • Alternative names

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

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-Firefly Luciferase antibody [EPR17790] (ab185924)
      Immunocytochemistry/ Immunofluorescence - Anti-Firefly Luciferase antibody [EPR17790] (ab185924)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK293T (Human epithelial cells from embryonic kidney) cells transfected with Empty vector or Firefly Luciferase, labeling Firefly Luciferase with ab185924 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on HEK293T cells transfected with Firefly Luciferase is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
      The negative controls are as follows;
      -ve control 1: ab185924 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

    • Flow Cytometry - Anti-Firefly Luciferase antibody [EPR17790] (ab185924)
      Flow Cytometry - Anti-Firefly Luciferase antibody [EPR17790] (ab185924)

      Flow Cytometry analysis of 293T (Human epithelial cell line from embryonic kidney) cells transfected with GFP tagged Firefly Luciferase labeled with purified ab185924 at 1/70 dilution (10µg/ml, Right panel). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 647)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Left panel) was used as the isotype control.

    • Western blot - Anti-Firefly Luciferase antibody [EPR17790] (ab185924)
      Western blot - Anti-Firefly Luciferase antibody [EPR17790] (ab185924)
      All lanes : Anti-Firefly Luciferase antibody [EPR17790] (ab185924) at 1/20000 dilution

      Lane 1 : Empty vector (vector control) transfected HEK293 (Human embryonic kidney) whole cell lysate
      Lane 2 : Firefly Luciferase transfected HEK293 (Human embryonic kidney) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

      Predicted band size: 61 kDa
      Observed band size: 61 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Firefly Luciferase antibody [EPR17790] (ab185924)
      Western blot - Anti-Firefly Luciferase antibody [EPR17790] (ab185924)
      All lanes : Anti-Firefly Luciferase antibody [EPR17790] (ab185924) at 1/1000 dilution

      Lane 1 : Firefly Luciferase transfected HEK293 (human embryonic kidney) whole cell lysate at 10 µg
      Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg
      Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
      Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg
      Lane 5 : SH-SY5Y (Human neuroblastoma from bone marrow cells) whole cell lysate at 20 µg
      Lane 6 : Human fetal brain lysate at 10 µg
      Lane 7 : Human fetal heart lysate at 10 µg
      Lane 8 : Human fetal kidney lysate at 10 µg
      Lane 9 : Human fetal spleen lysate at 10 µg

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 61 kDa
      Observed band size: 61 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-Firefly Luciferase antibody [EPR17790] (ab185924)
      Western blot - Anti-Firefly Luciferase antibody [EPR17790] (ab185924)
      All lanes : Anti-Firefly Luciferase antibody [EPR17790] (ab185924) at 1/1000 dilution

      Lane 1 : Firefly Luciferase transfected HEK293 (Human embryonic kidney) whole cell lysate
      Lane 2 : Mouse brain lysate
      Lane 3 : Mouse heart lysate
      Lane 4 : Mouse kidney lysate
      Lane 5 : Mouse spleen lysate
      Lane 6 : Rat brain lysate
      Lane 7 : Rat heart lysate
      Lane 8 : Rat kidney lysate
      Lane 9 : Rat spleen lysate
      Lane 10 : C6 (Rat glial tumor cells) whole cell lysate
      Lane 11 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
      Lane 12 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
      Lane 13 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

      Predicted band size: 61 kDa
      Observed band size: 61 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    References

    This product has been referenced in:

    • Ryu HG  et al. Upf1 regulates neurite outgrowth and branching by transcriptional and post-transcriptional modulation of Arc. J Cell Sci 132:N/A (2019). Read more (PubMed: 30584064) »
    • Benhabiles H  et al. Optimized approach for the identification of highly efficient correctors of nonsense mutations in human diseases. PLoS One 12:e0187930 (2017). Read more (PubMed: 29131862) »
    See all 4 Publications for this product

    Publishing research using ab185924? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-2 of 2 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Firefly Luciferase antibody [EPR17790]

     Below Average
    Abreviews
    This product is known to not work in this application or species.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Pancreas and Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Sodium Citrate
    Permeabilization
    No
    Specification
    Pancreas and Liver
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 12 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Firefly Luciferase antibody [EPR17790]

     Inconclusive
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Pancreas)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate (10mM)
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Pancreas
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 14 2016

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