Recombinant
RabMAb

Recombinant Anti-Firefly Luciferase antibody [EPR17789] - N-terminal (ab185923)

Overview

  • Product name

    Anti-Firefly Luciferase antibody [EPR17789] - N-terminal
    See all Firefly Luciferase primary antibodies
  • Description

    Rabbit monoclonal [EPR17789] to Firefly Luciferase - N-terminal
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, Flow Cytmore details
  • Species reactivity

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

    Synthetic peptide within Firefly Firefly Luciferase aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: P08659

  • Positive control

    • WB: Firefly Luciferase transfected HEK-293 whole cell lysate. ICC/IF: Firefly Luciferase transfected HEK-293T 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, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR17789
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab185923 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
ICC/IF 1/500.
WB 1/2000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
Flow Cyt 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
  • Database links

  • Alternative names

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

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cells from embryonic kidney) cells transfected with Empty vector or Firefly Luciferase, labeling Firefly Luciferase with ab185923 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: ab185923 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 analysis of 293T (Human epithelial cell line from embryonic kidney) transfected with GFP tagged Firefly Luciferase cells labeling Firefly Luciferase with purified ab185923 at 1/200 dilution (10ug/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.

  • All lanes : Anti-Firefly Luciferase antibody [EPR17789] - N-terminal (ab185923) at 1/2000 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 (HRP) (ab97051) at 1/100000 dilution

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


    Exposure time: 30 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Firefly Luciferase antibody [EPR17789] - N-terminal (ab185923) at 1/1000 dilution

    Lane 1 : Firefly Luciferase transfected HEK293 (human embryonic kidney) whole cell lysate at 10 µg
    Lane 2 : HeLa (human cervix adenocarcinoma) whole cell lysate at 20 µg
    Lane 3 : MCF-7 (human breast carcinoma) whole cell lysate at 20 µg
    Lane 4 : Jurkat (human acute T cell leukemia) whole cell lysate at 20 µg
    Lane 5 : SH-SY5Y (human neuroblastoma) whole cell lysate at 20 µg
    Lane 6 : Human fetal heart lysate at 10 µg
    Lane 7 : Human fetal kidney lysate at 10 µg
    Lane 8 : Human fetal spleen lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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


    Exposure time: 3 minutes


    Blocking and dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Firefly Luciferase antibody [EPR17789] - N-terminal (ab185923) 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 kidney lysate
    Lane 8 : Rat spleen lysate
    Lane 9 : C6 (rat glioma) whole cell lysate lysate
    Lane 10 : Raw264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate
    Lane 11 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 12 : NIH/3T3 (mouse embryo) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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


    Exposure time: 3 minutes


    Blocking and dilution buffer: 5% NFDM/TBST.

References

This product has been referenced in:

  • Azad T  et al. A LATS biosensor screen identifies VEGFR as a regulator of the Hippo pathway in angiogenesis. Nat Commun 9:1061 (2018). WB . Read more (PubMed: 29535383) »
  • Yan J  et al. Molecular characterization of the giant freshwater prawn (Macrobrachium rosenbergii) beta-actin gene promoter. PeerJ 6:e5701 (2018). Read more (PubMed: 30386688) »
See all 3 Publications for this product

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