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G5420 Sigma-Aldrich

Anti-β-Glucuronidase (N-Terminal) antibody produced in rabbit

~1.5 mg/mL, affinity isolated antibody, buffered aqueous solution

Synonym: Anti-GUS

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Properties

Related Categories Alphabetical Index, Antibodies, GD-GL, Plant Biology, Primary Antibodies More...
biological source   rabbit
antibody form   affinity isolated antibody
antibody product type   primary antibodies
clone   polyclonal
form   buffered aqueous solution
mol wt   antigen mol wt 60 kDa
species reactivity   plant
concentration   ~1.5 mg/mL
application(s)   western blot: 0.5-1 μg/mL using GUS purified from E. coli
conjugate   unconjugated
shipped in   dry ice
storage temp.   −20°C

Description

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.

Features and Benefits

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Immunogen

sythetic peptide corresponding to amino acids 1-16 located at the N-terminus of E. coli GUS, conjugated to KLH.

Specificity

Anti-β-Glucuronidase (N-Terminal) recognizes bacterial GUS expressed in transgenic tobacco plants.

Physical form

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

Application

Rabbit polyclonal anti-β-Glucuronidase (N-Terminal) antibody may be used for the detection of GUS by immunoblotting (60 kDa). It may be used to detect the reporter GUS marker protein in transgenic plants.

Biochem/physiol Actions

β-Glucuronidase catalyzes the cleavage of terminal glucuronic acid bound by β-linkage from mono-, oligo-, or polysaccharides or phenols. GUS is mainly used as a marker during transgenic events. In vitro GUS activity assays predominantly facilitates histochemical and fluorometric localization in tissues. GUS activity is also used for the analysis of expression of foreign genes. GUS reporter genes fused with specific promoter genes have been used for the determination of transformed tissues on culture media containing antibiotics. GUS-specific antibodies are helpful in detecting GUS gene product in transformed plants.

General description

Reporter genes are widely used for studying the expression of foreign genes in transformed plants tissues. Using appropriate promoter-reporter gene constructs, this technique allows an independent verification of the transformed status of tissues growing on media containing selective antibiotics or herbicides. In addition, it serves as a principal means to follow gene transfer and monitor genetic transformation of plant species. Several screenable markers are available including β-glucuronidase (GUS), β-galactosidase (β-GAL), chloramphenicol acetyl transferase (CAT), green fluorescent protein (GFP) and luciferase. E. coli GUS has been extensively used to monitor transgene delivery to plant tissue. Encoded by the E. coli gus gene (also referred to as uidA), GUS protein (60 kDa) is an hydrolase that catalyses the cleavage of a variety of b-glucuronide derivatives available for colorimetric, fluorimetric and histochemical assays. Several features make the gus gene superior as a reporter gene for plant studies and in the production of genetically engineered crops.
Many plants lack detectable endogenous glucuronidase activity, resulting in essentially no background. In addition, GUS activity is easily assayed in vitro and can withstand fixation, enabling histochemical localization in cells and tissue sections. However, one of the major limitations of the gus reporter gene system is that histochemical GUS assay system is destructive for the plant tissue, and therefore it is not suitable for direct visual selection of transformed plants. Antibodies specific for GUS are useful tools for detecting GUS gene product in transformed plants.

Storage and Stability

For continuous use, store at 2-8 °C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing, or storage in “frostfree” freezers, is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilutions should be discarded if not used within 12 hours.

Safety & Documentation

Safety Information

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

Articles

Antibody Basics

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Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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G5545 Anti-β-Glucuronidase (C-Terminal) antibody produced in rabbit, ~1.5 mg/mL, affinity isolated antibody, buffered aqueous solution

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