Luciferase Antibody [HRP]

• Reactivity: Pp
• Applications: WB, ELISA
• Clonality: Polyclonal
• Host: Goat
• Conjugate: HRP
• Concentration: LYOPH

Luciferase Antibody [HRP] Summary

• Description: Store vial at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.
  
  This product is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, anti-Goat Serum as well as purified and partially purified Luciferase [Photinus pyralis (Firefly)]
• Immunogen: Luciferase [Photinus pyralis (Firefly)]
• Specificity: No reactivity is observed against Sea pansy (Renilla reniformis) luciferase.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • ELISA 1:2000-1:30000
  • Western Blot 1:500-1:2000
• Application Notes: This product is produced from Photinus pyralis (Firefly) which produces a green light with a wavelength of 562 nm. Anti-Luciferase Peroxidase has been tested by ELISA and western blot. Expect a band at approximately 60.7 kDa in size corresponding to Luciferase by western blotting in the appropriate cell lysate or extract. Anti-Luciferase has been assayed against 1.0 ug of Luciferase [Photinus pyralis (Firefly)] in a standard capture ELISA using ABTS (2,2'-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) as a substrate for 30 minutes at room temperature. A working dilution of 1:50,000 to 1:200,000 of the reconstitution concentration is suggested for this product.
  
  NOTE: Do NOT add Sodium Azide (it inhibits HRP irreversibly).
• Readout System:
  • NovaLume Plus Chemiluminescent Reagent LUM-555

Reactivity Notes

No reactivity is observed against Sea pansy (Renilla reniformis) luciferase.

Packaging, Storage & Formulations

• Storage: Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.
• Buffer: Lyophilized from 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
• Preservative: 0.01% Gentamicin Sulfate
• Concentration: LYOPH
• Purity: Multi-step
• Reconstitution Instructions: Reconstitute with 100 ul deionized water (or equivalent)

Alternate Names for Luciferase Antibody [HRP]

• 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.

Publications for Luciferase Antibody (NB100-1676)(1)

Publication using NB100-1676

• Yang G, Abate A, George AG, et al. Maturational differences in lung NF-kappaB activation and their role in tolerance to hyperoxia J Clin Invest 2004-09-03 [PMID: 15343385]