Luciferase (firefly) Antibody - BSA Free

Western Blot: Luciferase (firefly) Antibody [NBP3-33161] - Western Blot of Luciferase (firefly) Antibody. Lane 1: Luciferase (Firefly). Load: 50 ng per lane. Primary antibody: Luciferase (firefly) Antibody at 1:1,000 overnight at 4°C. Secondary antibody: HRP goat secondary antibody at 1:40,000 for 30 min at RT. Block for 30 min at RT. Predicted/Observed size: 60 kDa, 60 kDa for Luciferase.

• Reactivity: Pp
• Applications: WB
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: LYOPH

Luciferase (firefly) Antibody - BSA Free Summary

• Description: This product was prepared from monospecific antiserum by a delipidation and defibrination. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-goat serum, purified and partially purified Luciferase [Firefly].
  
  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.
• Immunogen: Luciferase [Firefly] Native Protein
• Specificity: No reactivity is observed against Sea pansy (Renilla reniformis) luciferase.
• Clonality: Polyclonal
• Host: Goat
• Purity: Delipidation and Defibrination

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This has been tested by western blot and is suitable in ELISA. This product can be assayed against 1.0 ug of Luciferase [Firefly] in a standard ELISA using Peroxidase conjugated Affinity Purified anti-Goat IgG [H&L] (Goat) and (ABTS (2,2'-azino-bis-[3-ethylbenthiazoline-6-sulfonic acid]) as a substrate for 30 minutes at room temperature. A working dilution of 1:20,000 to 1:100,000 of the reconstitution concentration is suggested for this product.

Packaging, Storage & Formulations

• Storage: Store at 4C. After reconstitution store at -20C.
• Buffer: Lyophilized from 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Concentration: LYOPH
• Purity: Delipidation and Defibrination
• Reconstitution Instructions: Reconstitute with 2.0 ml with deionized water (or equivalent).

Alternate Names for Luciferase (firefly) Antibody - BSA Free

• firefly luciferase

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.