• USA Home
  • L0159 - Anti-Luciferase antibody produced in rabbit

EMAIL THIS PAGE TO A FRIEND
L0159 Sigma

Anti-Luciferase antibody produced in rabbit

IgG fraction of antiserum, buffered aqueous solution

Purchase

Properties

Related Categories Alphabetical Index, Analytical and Industrial Enzymes, Antibodies, Biochemicals and Reagents, Enzymes, Inhibitors, and Substrates,
biological source   rabbit
antibody form   IgG fraction of antiserum
clone   polyclonal
form   buffered aqueous solution
application(s)   indirect immunofluorescence: 1:500-1:1,000 using eukaryotic cells transfected with a plasmid bearing the luciferase gene
  western blot: 1:10,000 using purified Luciferase
conjugate   unconjugated
shipped in   dry ice
storage temp.   −20°C

Description

Immunogen

firefly (Photinus pyralis) luciferase.

Specificity

The antibody identifies recombinant luciferase in eukaryotic cells transfected with a plasmid bearing the luciferase gene.

Physical form

Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.

Application

Anti-Luciferase antibody used to detect luciferase can provide an alternative detection assay. This assay directly detects luciferase protein levels, and thus has the advantage that it does not require luciferase activity and is not dependent on rapid kinetics. Moreover, antibodies can detect the luciferase enzyme expression in situ, providing a means to study the localized signal sequences using luciferase as a reporter gene.
A working concentration of 10 mg/ml is obtained on methanol-acetone fixed transfected* cells using an FITC conjugated secondary antibody in an immunofluorescence assay.

General description

Analysis of gene expression is commonly assayed by transient transfection. These systems are usually based on the use of fusion genes which are inserted into cells, and expression of the gene is assayed within 48 hours after introduction of DNA. Usually the fusion consists of the promoter binding site or enhancer sequence under study which is attached to a reporter gene. The amount of the reporter protein synthesized under the experimental conditions is presumed to reflect the ability of the sequences studied to direct or promote transcription.
Luciferase has become one of the more widely used reporter enzymes. The reporter plasmid contains the gene from the firefly Photinus pyralis. The enzyme catalyzes a bioluminescent reaction which requires the substrate luciferin as well as Mg+2 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. The luciferase assay is fast and sensitive and does not require a radioactive substrate as in the CAT assay. A disadvantage of the luciferase assay is that it requires a rather expensive instrument, the luminometer, to measure enzyme activity.

Price and Availability


Amplified Detection
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Related Content

Antibody Explorer - Search 80,000 Antibodies at Sigma-Aldrich

Monoclonal and polyclonal primary antibodies are focused on cell biology, neurobiology and molecular biology. Secondary antibodies targeting multiple host’s IgG are conjugated to alkaline phosphatase...
Keywords: Amplification, Buffers, Cell biology, Enzyme-linked immunosorbent assay, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Molecular biology, Phosphorylations, Purification, Western blot

Peer-Reviewed Papers
15

References

Set your institution to view full text papers.
Related Products

related product

Product #

Description

Add to Cart

06693 Timestrip Plus -20 °C

Technical Service:

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Bulk Ordering & Pricing:

Need larger quantities for your development, manufacturing or research applications?