Key features and details
- Mouse monoclonal [Luci17] to Firefly Luciferase
- Suitable for: WB
- 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
Fusion protein corresponding to Firefly Firefly Luciferase.
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.
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
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.
|WB||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.
- ec 22.214.171.124 antibody
- Firefly antibody
- Luciferase antibody
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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 14 publications.
- 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
- Forootan SS et al. Real-time in vivo imaging reveals localised Nrf2 stress responses associated with direct and metabolism-dependent drug toxicity. Sci Rep 7:16084 (2017). PubMed: 29167567
- Davis MI et al. Small Molecule Inhibition of the Ubiquitin-specific Protease USP2 Accelerates cyclin D1 Degradation and Leads to Cell Cycle Arrest in Colorectal Cancer and Mantle Cell Lymphoma Models. J Biol Chem 291:24628-24640 (2016). PubMed: 27681596
- Kao T et al. GAPTrap: A Simple Expression System for Pluripotent Stem Cells and Their Derivatives. Stem Cell Reports 7:518-26 (2016). WB, Flow Cyt . PubMed: 27594589