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

11-20 of 35 Abreviews or Q&A

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

Answer

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 to find one in a publication:

https://www.abcam.com/Firefly-Luciferase-antibody-ab21176-references.html

I hope this information helps. Please contact us with any other questions.

Read More

Answer

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 assistance with troubleshooting or a credit, as this product is guaranteed for IF under our Abpromise. Please provide some additional protocol information.


Regarding using the antibody in WB, we are not sure if this antibody will be suitable for western, but would recommend a starting dilution of 1 ug/ml.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or concerns.

Read More

Answer

Thank you for contacting us.

The customer could use lysates of HEK-293T cells overexpressing luciferase as positive control for ab21176.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Thank you for taking time to send me these details and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.

Having reviewed the protocol, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial.

I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More
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

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

Read More

Question
Answer

Thank you for your enquiry.

Here is an example of a secondary antibody you can use to detect ab64569:

https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-Fc-DyLight-488-pre-adsorbed-ab98712.html

ab98712 is conjugated to a fluor that can be visualized with the same filter set as is used for FITC. This antibody has been preadsorbed to rat serum (among others) and has shown minimum cross reactivity to rat immunoglobulins.

We have a couple useful tools on our website for selecting secondary antibodies. One is a search and another is a browse feature and both can be found at:
https://www.abcam.com/index.html?pageconfig=productmap&cl=918

I hope this is helpful. Please contact me again if you have any further questions.

Read More

11-20 of 35 Abreviews or Q&A

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