ANTI-RENILLA LUCIFERASE CLONE 1D5.2 (C005B-452220)

General description

Coelenterazine h 2-monooxygenase (UniProt: P27652; also known as EC:1.13.12.5, Renilla-luciferin 2-monooxygenase, Renilla-type luciferase) is a monomeric, bioluminescent protein derived from the sea pansy (Renilla reniformis). Due to its light emitting abilities, the luciferase from Renilla reniformis has been widely used in molecular biology as a reporter gene in cell culture experiments and small animal imaging. Renilla luciferase is shown to be present in membrane-bound intracellular structures within specialized light-emitting cells along with a closely interacting green fluorescent protein (RrGFP) and a calcium- activated luciferin-binding protein. Upon binding the substrate, it catalyzes an oxygenation reaction, producing a very short-lived hydroperoxide that cyclizes into a dioxetanone structure that collapses, releasing a CO2 molecule. The spontaneous breakdown of the dioxetanone releases the energy that is necessary to generate the excited state of the coelenteramide product, which is the singlet form of the monoanion. The optimum pH for this reaction is 7.4. It is shown that under in vivo conditions the product undergoes the process of nonradiative energy transfer to an accessory protein, a GFP, which results in green bioluminescence (~ 509 nm). However, in vitro, in the absence of GFP, it emits blue light (~480 nm). (Ref.: Loening, AM., et al. (2007). J. Mol. Biol. 374(4); 1017-1028; Lorenz, WW., et al. (1991). Proc. Natl. Acad. Sci. USA 88(10); 4438-4442).

Specificity

Clone 5B11.2 is a mouse monoclonal antibody that detects Renilla-type luciferase (Renilla-luciferin 2-monooxygenase).

Immunogen

GST-tagged full-length recombinant Renilla reniformis Luciferase.

Application

Quality Control Testing

Evaluated by ELISA with recombinant Renilla reniformis Luciferase.

ELISA Analysis (ELISA): Various dilution (starting at 5 μg/mL, two-fold serial dilutions; 8 points) of this antibody detected recombinant Renilla reniformis luciferase.

Tested Applications

Immunoprecipitation Analysis: A representative lot immunoprecipitated Renilla Luciferase in Immunoprecipitation applications (Adelfinger, L., et. al. (2014). Biochem Pharmacol. 91(3):369-79).

Western Blotting Analysis: A representative lot detected Renilla Luciferase in Western Blotting applications (Simms, C.L., et. al. (2019). Cell Rep. 28(7):1679-1689; Lu, Q., et. al. (2018). Nat Commun. 9(1):3104).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Anti-Renilla Luciferase, clone 1D5.2, Cat. No. MAB4410-I, is a mouse monoclonal antibody that detects Renilla-type luciferase (Renilla-luciferin 2-monooxygenase) and is tested for use in ELISA, Immunoprecipitation, and Western Blotting.

Physical form

Purified mouse monoclonal antibody IgG in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at +2°C to +8°C from date of receipt.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.