Product nameAnti-Firefly Luciferase antibody [Luci17]
See all Firefly Luciferase primary antibodies
DescriptionMouse monoclonal [Luci17] to Firefly Luciferase
Tested applicationsSuitable for: Flow Cyt, WB, IHC-Frmore details
Hybridoma produced by the fusion of spleenocytes from mice immunized with luciferase protein isolated from Photinus pyralis.
- WB: Purified luciferase protein. IHC: frozen drosophila embryos
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.08% Sodium Azide
Constituents: PBS, pH 7.2
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.
|Flow Cyt||Use at an assay dependent concentration. PubMed: 18612415
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use at an assay dependent concentration.|
|IHC-Fr||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 220.127.116.11 antibody
- Firefly antibody
- Luciferase antibody
- Luciferin 4 monooxygenase antibody
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.
Flow cytometric analysis of HEK293 cells expressing Firefly Luciferase, staining Firefly Luciferase with ab16466.
Cells were fixed with paraformaldehyde at room temperature for 20 min, rinsed twice with permeabilization buffer (PBS, 1% FBS, 0.1% saponin), and incubated with primary antibody. A FITC-conjugated anti mouse IgG was used as the secondary antibody.
This product has been referenced in:
- 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). Read more (PubMed: 29167567) »
- 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 . Read more (PubMed: 27594589) »