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

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1-10 of 17 Abreviews

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

Abcam user community

Verified customer

Submitted Nov 12 2013

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

Abcam user community

Verified customer

Submitted Mar 23 2012

Application
Flow Cytometry
Sample
Human Cell (Jurkat)
Permeabilization
Yes - eBio 1x Perm Buffer Foxp3 kit, Methanol, Acetone
Gating Strategy
cells, singlets
Specification
Jurkat
Fixation
eBio Fix/Perm, 2%FA, Acetone

Abcam user community

Verified customer

Submitted Mar 06 2019

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse neural stem cell)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
40 µg
Treatment
expressing firefly luciferase
Specification
mouse neural stem cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jun 12 2018

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

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

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

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

Dr. Francois Daubeuf

Verified customer

Submitted Dec 05 2016

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

Abcam user community

Verified customer

Submitted Dec 31 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (mammary gland)
Specification
mammary gland
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate based, microwave heated
Permeabilization
Yes - triton-X 0.25%
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C

Dr. yael raz

Verified customer

Submitted Jun 07 2012

1-10 of 17 Abreviews

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