Luciferase Antibody - BSA Free

Western Blot: Luciferase Antibody [NB110-17348] - Lysates of HEK-293T cells overexpressing luciferase was separated on SDS-PAGE and probed with Anti-Luciferase The antibody was developed using Goat Anti-Rabbit IgG-Alkaline Phosphatase and a colorimetric substrate. Lanes 1. Antibody dilution 1:1,000 2. Antibody dilution 1:1,000 + luciferase immunizing peptide

Immunocytochemistry/Immunofluorescence: Luciferase Antibody [NB110-17348] - HEK-293T cells overexpressing luciferase were fixed and stained 10 ug/mL Anti-Luciferase. The antibody was developed using Goat Anti-Rabbit IgG, FITC conjugate.

• Reactivity: Pp
• Applications: WB, ICC/IF
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

Luciferase Antibody - BSA Free Summary

• Description: Novus Biologicals Rabbit Luciferase Antibody - BSA Free (NB110-17348) is a polyclonal antibody validated for use in WB and ICC/IF. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: This Luciferase Antibody was developed against full length native protein (purified) (Firefly (Photinus pyralis)).
• Localization: Peroxisomal
• Specificity: This Luciferase Antibody identifies recombinant luciferase in eukaryotic cells transfected with a plasmid bearing the luciferase gene.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Purity: IgG purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:500-1:1000
  • Western Blot 1:10000
• Application Notes: ICC/IF: A working dilution of 1:5001:1,000 is obtained on methanol-acetone fixed transfected cells using an FITC conjugated secondary antibody. Cells were transfected with a reporter plasmid containing the gene for luciferase.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 10mM PBS (pH 7.4)
• Preservative: 0.09% Sodium Azide
• Purity: IgG purified

Alternate Names for Luciferase Antibody - BSA Free

• ec 1.13.12.7
• firefly luciferase
• Fluc
• LuC
• Luciferase
• luciferin 4 monooxygenase
• Luciferin 4-monooxygenase

Background

Luciferase is a generic term for a group of oxidative enzymes used in bioluminescence. Firefly (Photinus pyralis) and bacterial luciferase enzymes are commonly used in assay systems such as cell viability assays, reporter gene assays, and for in vivo imaging. Bacterial luciferases are flavoenzymes composed of two subunits each encoded by the luxA and luxB genes, while the firefly luciferase is a single polypeptide specified by the luc gene (1). Firefly luciferase (theoretical molecular weight: 61 kDa) oxidizes the substrate luciferin to oxyluciferin in a bioluminescent reaction requiring Mg2+ and ATP (2,3). This reaction produces a flash of yellow-green light with an emission peak around 560nm that can be detected by a luminometer (3). Firefly luciferase has become one of the more widely used reporter proteins and is an excellent tool for the study of gene expression, given that the amount of light emitted is directly proportional to luciferase activity (4).

The luciferase assay is fast and sensitive, differentiating itself from the CAT (chloramphenicol acetyltransferase) assay because it does not require a radioactive substrate.

References

1. Eun, H. (1996). Marker/Reporter enzymes. Enzymology Primer for Recombinant DNA Technology, 567-645. doi:10.1016/b978-012243740-3/50011-9

2. McNabb, D. S., Reed, R., & Marciniak, R. A. (2005). Dual luciferase assay system for rapid assessment of gene expression in Saccharomyces cerevisiae. Eukaryotic Cell, 4(9), 1539-1549. doi:10.1128/ec.4.9.1539-1549.2005

3. Fraga, H. (2008). Firefly luminescence: A historical perspective and recent developments. Photochemical & Photobiological Sciences, 7(2), 146-158. doi:10.1039/b719181b

4. Younes, A., Lukyanenko, Y. O., Lyashkov, A. E., Lakatta, E. G., & Sollott, S. J. (2011). A bioluminescence method for direct measurement of phosphodiesterase activity. Analytical Biochemistry, 417(1), 36-40. doi:10.1016/j.ab.2011.05.036

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Review for Luciferase Antibody (NB110-17348) (1)

Average Rating: 1

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Verified Customer Immunofluorescence-Frozen Mouse 01/31/2024

Summary

• Application: Other
• Sample Tested: Mouse spleen, mouse thymus,Thymus tissue
• Species: Mouse
• Lot: 049M4778V

Comments

• Comments: ***Bio-Techne Response: This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.*** Frozen Sections were fixed for 10 min in 4% PFA at room temp followed by incubation in 100% Methanol at -20 C for 10 min. Sections were washed and blocked using 10% Donkey Serum or 2% Fish Gelatin/10% Donkey Serum. Tested Primary Dilutions: 1:500, 1:1000, and 1:5000 overnight at 4 C. Secondary Tested: Preabsorbed Anti-Rabbit 488 or Rabbit 555. Tissue collected from transgenic mouse expressing luciferase from CAG promoter. Luciferase activity was confirmed by measuring luminescence from collected tissue before embedding in OCT.