Recombinant
RabMAb

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

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

  • 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 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.

  • 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.

  • 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.

  • 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:

  • 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) »
  • Li D  et al. An Ebola Virus-Like Particle-Based Reporter System Enables Evaluation of Antiviral Drugs In Vivo under Non-Biosafety Level 4 Conditions. J Virol 90:8720-8 (2016). Read more (PubMed: 27440895) »
See all 3 Publications for this product

Customer reviews and Q&As

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1-2 of 2 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

Abcam user community

Verified customer

Submitted Aug 12 2016

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)
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

Abcam user community

Verified customer

Submitted Jul 14 2016

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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