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:
  • Revaud J  et al. Firewalls Prevent Systemic Dissemination of Vectors Derived from Human Adenovirus Type 5 and Suppress Production of Transgene-Encoded Antigen in a Murine Model of Oral Vaccination. Front Cell Infect Microbiol 8:6 (2018). Read more (PubMed: 29423380) »
  • Zheng J  et al. Evaluation of metastatic niches in distant organs after surgical removal of tumor-bearing lymph nodes. BMC Cancer 18:608 (2018). Read more (PubMed: 29848296) »
See all 38 Publications for this product

Customer reviews and Q&As

1-10 of 35 Abreviews or Q&A

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

Answer

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 included.

I hope that this helps and please do not hesitate to contact us if you require any further information or assistance.

Read More

Answer

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 of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Read More

Answer

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 using the product in applications listed on the datasheet and contact us within six months of purchase, we are also happy to replace or refund the product should we not be able to help you to resolve the issue.

Would you be able to send me the details of your protocol so that I may get a better understanding of the problemms which you are facing. These details are also important should we decide that this product needs further internal testing.

Could you also provide the Abcam order confirmation number or PO used?



Thank you for your cooperation, I look forward to your response. 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
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

Abcam user community

Verified customer

Submitted Mar 23 2012

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: low Target retrival solution
Permeabilization
No
Specification
Liver
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Frau. Ines Kleiber-Schaaf

Verified customer

Submitted Nov 23 2018

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

1-10 of 35 Abreviews or Q&A

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