Overview

  • Product name
    Anti-Firefly Luciferase antibody
    See all Firefly Luciferase primary antibodies
  • Description
    Rabbit polyclonal to Firefly Luciferase
  • Specificity
    The antibody identifies by immunocyotochemistry recombinant luciferase in cultured HEK293 cells transfected with a plasmid bearing the luciferase gene.
  • Tested applications
    Suitable for: ICC/IF, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Firefly
  • Immunogen

    Full length native protein (purified) (Firefly (Photinus pyralis)).

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 15mM Sodium Azide
    Constituents: 0.01M PBS, pH 7.4
  • Concentration information loading...
  • Purity
    IgG fraction
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab21176 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 Use a concentration of 10 µg/ml.
IHC-Fr 1/1000. PubMed: 18219389
WB 1/1000 - 1/2000.

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

Anti-Firefly Luciferase antibody images

  • ab21176 at 10 - 20  µg/mL staining Luciferase in transfected HEK293 cells by ICC/IF. The cells were fixed with methanol and acetone. A FITC conjugated Anti-Rabbit IgG was used as the secondary antibody. 

  • ab21176 at 1/200 dilution staining engineered adult rat stromal stem cells by ICC/IF. The cells were paraformaldehyde (2%) fixed and Triton X-100 (0.1%) was used for cell permealization (15 min incubation time). The cells were incubated with the antibody overnight at 4C. The image shows Luciferase (green-upper right panel), counterstained cell nuclei (DAPI-blue-upper left panel), overlay (lower left panel) and a phase contrast image (lower right panel). The image was taken with a confocal laser scanning microscope equipped with an additional laser differential Interference Contrast (DIC) mode.

    See Abreview

  • All lanes : Anti-Firefly Luciferase antibody (ab21176) at 1/1000 dilution

    Lane 1 : Lysates of HEK-293T cells overexpressing luciferase
    Lane 2 : Lysates of HEK-293T cells overexpressing luciferase with Luciferase Immunizing Peptide

    Secondary
    Goat Anti-Rabbit IgG-Alkaline Phosphatase and a colorimetric substrate
  • ab21176 staining Firefly Luciferase in Mouse cerebellum tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone, permeabilized with PBS + 0.1% Triton X-100 (PBST) for 10 minutes and blocked with 10% serum for 1 hour at 22°C. Samples were incubated with primary antibody (1/1500 in 10% Goat serum in PBST) for 1 hour at 22°C. An Alexa Fluor® 555-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • ab21176 staining mouse mammary carcinoma cells by ICC/IF.  Cells were PFA fixed and permeabilized in 0.1% Triton X-100 prior to blocking with a commercial blocking agent.  The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 25°C.  An Alexa Fluor® 555 conjugated goat anti-rabbit antibody was used as the secondary.  The image shows firefly luciferase (red) in mouse tumour cells and cell nuclei counterstained with Hoechst (blue).

    See Abreview

  • Immunocytochemical analysis analysis of HEK-293T cells overexpressing luciferase labelling Luciferase with ab21176 at a concentration of 10μg/mL. The secondary antibody was a Goat Anti-Rabbit IgG, FITC conjugate.

References for Anti-Firefly Luciferase antibody (ab21176)

This product has been referenced in:
  • Parkins KM  et al. A multimodality imaging model to track viable breast cancer cells from single arrest to metastasis in the mouse brain. Sci Rep 6:35889 (2016). IHC ; Mouse . Read more (PubMed: 27767185) »
  • Hayashi M  et al. Whole-Body In Vivo Monitoring of Inflammatory Diseases Exploiting Human Interleukin 6-Luciferase Transgenic Mice. Mol Cell Biol 35:3590-601 (2015). IHC . Read more (PubMed: 26283726) »

See all 14 Publications for this product

Product Wall

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (5-20% gradient gel)
Sample
Human Cell lysate - whole cell (293T)
Specification
293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Nov 12 2013

Thank you for your inquiry and your patience with this reply.

After some further digging, the lab was able to provide a PPT of the ICC/IF results of untransfected vs transfected HEK293 cells stained using ab21176. The protocol is also include...

Read More

Thank you for contacting us. Your credit note ID is xxxx. I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice o...

Read More

Thank you for contacting us.





I am sorry that this product has not performed as should be expected. This product is covered by our Abpromise guarantee. We are happy provide scientific support at any time. If you are us...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Panc1 cancer cell xenografts)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
Yes - TBST
Specification
Panc1 cancer cell xenografts
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 23 2012

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate pH6.0
Permeabilization
Yes - 0.25%Triton X100 in PBS
Specification
Tumor
Blocking step
Dakocytomation X0909 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
Fixative
Formaldehyde
Username

Hongwei Shao

Verified customer

Submitted Aug 28 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing (4-12 Tris-Bis)
Loading amount
40 µg
Specification
HEK293
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted May 31 2017

Application
Flow Cytometry
Sample
Human Cell (breast cancer cell line transfected to express luc)
Permeabilization
Yes - Cells were permeabilized using commercial kit
Gating Strategy
DAPI+ cells
Specification
breast cancer cell line transfected to express luc
Preparation
Cell harvesting/tissue preparation method: Cells in culture
Sample buffer: PBS-EDTA 3mM
Fixation
Cells were fixed using commercial kit
Username

Dr. Francois Daubeuf

Verified customer

Submitted Dec 05 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (breast cancer cell line transfected to express luc)
Permeabilization
Yes - PBS-Triton x100 0.5% 10min
Specification
breast cancer cell line transfected to express luc
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Cytospined cells
Username

Dr. Francois Daubeuf

Verified customer

Submitted Dec 05 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (breast tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
Yes - tbst
Specification
breast tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 25°C
Fixative
formalin
Username

Abcam user community

Verified customer

Submitted Dec 31 2014

1-10 of 33 Abreviews or Q&A

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

Sign up