Key features and details
- Mouse monoclonal [Luci17] to Firefly Luciferase
- Suitable for: WB
- Reacts with: Mouse, Human
- Isotype: IgG1
Product nameAnti-Firefly Luciferase antibody [Luci17]
See all Firefly Luciferase primary antibodies
DescriptionMouse monoclonal [Luci17] to Firefly Luciferase
Tested applicationsSuitable for: WBmore details
Species reactivityReacts with: Mouse, Human
Recombinant fragment corresponding to Firefly Firefly Luciferase. Hybridoma produced by the fusion of splenocytes from mice immunized with luciferase protein isolated from Photinus pyralis.
Database link: P08659
- WB: Purified luciferase protein
Analysis of gene expression is most commonly assayed by transient transfection. Systems are generally based on the use of fusion genes which are inserted into cells, and the gene expression is assayed within 48 hours after introduction of DNA. Usually the fusion consists of the promoter binding site or enhancer sequence under study which is attached to a reporter gene. The amount of the reporter protein synthesized under the experimental conditions, is presumed to reflect the ability of the sequences studied to direct or promote transcription. Several enzymes are commonly used as reporter proteins, among them are chloramphenicol acetyl transferase (CAT), -galactosidase, human growth hormone (hGH) and luciferase. Luciferase has become one of the widely used reporter enzymes. The enzyme catalyzes a bioluminescent reaction which requires the substrate luciferin as well as Mg+2 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. The use of an antibody to detect luciferase can provide an alternative detection assay which directly detects luciferase protein levels, and thus has the advantage that it does not require luciferase activity and is not dependent on rapid kinetics. Moreover, antibodies can detect the luciferase enzyme expression in situ, providing a means to study the localized signal sequences using luciferase as a reporter gene. Reacts with Luciferase (Firefly) Protein.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.08% Sodium azide
Concentration information loading...
PurityProtein A/G purified
Light chain typeunknown
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab16466 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use at an assay dependent concentration.
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.
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All lanes : Anti-Firefly Luciferase antibody [Luci17] (ab16466) at 1 µg/ml
Lane 1 : Non-transfected 293 whole cell lysate
Lane 2 : Firefly Luciferase transfected 293 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : NIH3T3 whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/10000 dilution
Performed under reducing conditions.
Observed band size: 65 kDa why is the actual band size different from the predicted?
Lanes 1 - 4: Merged signal (red and green). Green – ab16466 observed at 65 kDa. Red - loading control, ab181602, observed at 37 kDa.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab16466 and ab181602 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at a 1:10000 dilution for 1hr at room temperature and then imaged.
ab16466 has been referenced in 17 publications.
- Cheng K et al. Bioengineered bacteria-derived outer membrane vesicles as a versatile antigen display platform for tumor vaccination via Plug-and-Display technology. Nat Commun 12:2041 (2021). PubMed: 33824314
- Afzal S et al. Anti-stress, Glial- and Neuro-differentiation Potential of Resveratrol: Characterization by Cellular, Biochemical and Imaging Assays. Nutrients 12:N/A (2020). PubMed: 32121454
- Hammill JA et al. A Cross-Reactive Small Protein Binding Domain Provides a Model to Study Off-Tumor CAR-T Cell Toxicity. Mol Ther Oncolytics 17:278-292 (2020). PubMed: 32368616
- Afzal S et al. Rat Glioma Cell-Based Functional Characterization of Anti-Stress and Protein Deaggregation Activities in the Marine Carotenoids, Astaxanthin and Fucoxanthin. Mar Drugs 17:N/A (2019). PubMed: 30909572
- Godfrey TC et al. The microRNA-23a cluster regulates the developmental HoxA cluster function during osteoblast differentiation. J Biol Chem 293:17646-17660 (2018). PubMed: 30242124