Overview

  • Product name

    Anti-Firefly Luciferase antibody
    See all Firefly Luciferase primary antibodies
  • Description

    Rabbit polyclonal to Firefly Luciferase
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-Fr, WBmore details
  • Species reactivity

    Reacts with: Firefly
  • Immunogen

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

  • Positive control

    • ICC/IF: Firefly Luciferase transfected HEK-293 cells. Adult rat stromal cells. HEK-293 cells. Mouse mammary carcinoma cells. WB: Lysates from HEK-293T cells overexpressing luciferase. IHC-Fr: Mouse cerebellum tissue.

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

    pH: 7.40
    Preservative: 0.097% Sodium azide
    Constituent: PBS
  • 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

Images

  • ab21176 at 10 µg/mL staining Luciferase in transfected HEK-293 (Human epithelial cell line from embryonic kidney) cells by ICC/IF.

    The cells were fixed with methanol and acetone. An FITC conjugated anti-Rabbit IgG was used as the secondary antibody. 

    Left panel: Un-transfected cells.

    Right panel: Transfected cells.

  • ab21176 at 1/200 dilution staining engineered adult rat stromal stem cells by ICC/IF.

    The cells were fixed in 2% paraformaldehyde and 0.1% Triton X-100 was used for cell permeabilization (15 minutes incubation time). The cells were incubated with the antibody overnight at 4°C. 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

  • 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

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

    Lane 1 : Lysates from HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells overexpressing luciferase
    Lane 2 : Lysates from HEK-293T cells overexpressing luciferase with Luciferase Immunizing Peptide

    Secondary
    All lanes : Goat Anti-Rabbit IgG-Alkaline Phosphatase and a colorimetric substrate
  • Immunocytochemical analysis analysis of HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells overexpressing luciferase labeling Luciferase with ab21176 at a concentration of 10 μg/mL. The secondary antibody was a Goat anti-Rabbit IgG, FITC conjugate.

  • 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

References

This product has been referenced in:

  • Fritzell K  et al. Sensitive ADAR editing reporter in cancer cells enables high-throughput screening of small molecule libraries. Nucleic Acids Res 47:e22 (2019). Read more (PubMed: 30590609) »
  • Kim S  et al. Optimizing live-animal bioluminescence imaging prediction of tumor burden in human prostate cancer xenograft models in SCID-NSG mice. Prostate 79:949-960 (2019). Read more (PubMed: 30958914) »
See all 45 Publications for this product

Customer reviews and Q&As

31-35 of 35 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse mammary carcinoma)
Specification
Mouse mammary carcinoma
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100
Blocking step
Perkin Elmer TNB as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 29 2008

Application
Immunocytochemistry/ Immunofluorescence
Sample
Hamster Cell (BHK)
Specification
BHK
Fixative
Methanol

Mr. Vladimir Kocoski

Verified customer

Submitted Nov 06 2006

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Engineered adult rat stromal stem cells)
Specification
Engineered adult rat stromal stem cells
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2%

Dr. Mal Niladri

Verified customer

Submitted Oct 09 2006

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Sample
Mouse Tissue sections (mouse cerebellum)
Specification
mouse cerebellum
Permeabilization
Yes - PBS + 0.1% tritonx100 (PBST) for 10 min
Fixative
Acetone

Ms. Radhi Praba Velayutham

Verified customer

Submitted Oct 08 2013

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Panc1 cancer cell xenografts)
Specification
Panc1 cancer cell xenografts
Fixative
Acetone
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Mar 22 2012

31-35 of 35 Abreviews or Q&A

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