Overview

  • Product nameAnti-Firefly Luciferase antibody
    See all Firefly Luciferase primary antibodies
  • Description
    Rabbit polyclonal to Firefly Luciferase
  • SpecificityThe antibody identifies by immunocyotochemistry recombinant luciferase in cultured HEK293 cells transfected with a plasmid bearing the luciferase gene.
  • Tested applicationsSuitable for: ICC/IF, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Firefly (Photinus pyralis)
  • Immunogen

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

Properties

  • FormLiquid
  • Storage instructionsShipped 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 bufferPreservative: 15mM Sodium Azide
    Constituents: 0.01M PBS, pH 7.4
  • Concentration information loading...
  • PurityIgG fraction
  • ClonalityPolyclonal
  • IsotypeIgG
  • 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

  • RelevanceLuciferase 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 localizationPeroxisome
  • Database links
    • Alternative names
      • ec 1.13.12.7 antibody
      • Firefly antibody
      • Luciferase antibody
      • Luciferin 4 monooxygenase antibody

    Anti-Firefly Luciferase antibody images

    • 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 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

    • 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:
    • Hegde V  et al. In vivo gene silencing following non-invasive siRNA delivery into the skin using a novel topical formulation. J Control Release 196:355-62 (2014). IHC-P ; Mouse . Read more (PubMed: 25449884) »
    • Rocca CJ  et al. rAAV9 combined with renal vein injection is optimal for kidney-targeted gene delivery: conclusion of a comparative study. Gene Ther 21:618-28 (2014). IHC-Fr . Read more (PubMed: 24784447) »

    See all 12 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...

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    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...

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    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...

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    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 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

    Thank you for your inquiry.

    I heard back from the testing lab that they did not have any cell staining data for ab21176 with untransfected cells.

    I can point you to our references section of the online datasheet as you may be able t...

    Read More

    Thank you for contacting Abcam regarding ab21176.


    I am sorry that you were experiencing difficulties with this antibody in IHC. If you believe this is due to the antibody being defective, I am happy to provide additional technical assista...

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    1-10 of 31 Abreviews or Q&A

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