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N2662 Sigma-Aldrich

Anti-Nucleolin antibody produced in rabbit

affinity isolated antibody, buffered aqueous solution

Synonym: Anti-C23

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Properties

Related Categories Alphabetical Index, Antibodies, Antibodies for Cell Biology, Antibodies for Epigenetics and Nuclear Signaling, Antibodies for Gene Regulation,
conjugate   unconjugated
clone   polyclonal
biological source   rabbit
application(s)   immunoprecipitation (IP): 1-2 μg using 250 μg RIPA lysate of human HeLa cells
  indirect immunofluorescence: 10-20 μg/mL using human HeLa cells
  western blot: 0.5-1.0 μg/mL using whole extract of mouse NIH3T3 cells or rat PC12 cells
species reactivity   mouse, human, rat
mol wt   antigen 105 kDa
form   buffered aqueous solution
shipped in   dry ice
storage temp.   −20°C
antibody form   affinity isolated antibody
antibody product type   primary antibodies
UniProt accession no.   P19338
Gene Information   human ... NCL(4691)
mouse ... Ncl(17975)
rat ... Ncl(25135)

Description

General description

Anti-Nucleolin is produced in rabbit using as immunogen a synthetic peptide of human nucleolin with C-terminal added cysteine conjugated to KLH. Nucleolin contains a specific bipartite nuclear localization signal sequence and three different structural domains. The N-terminal domain comprises of highly acidic regions separated by basic sequences and contains multiple phosphorylation sites. The central domain contains four RNA-binding domains (RBD or RRM) and the C-terminal domain (GAR or RGG) is rich in glycine, arginine, and phenylalanine residues. N and C terminal domains are involved in protein-protein interaction whereas the central domain is involved in specific interactions with nucleic acids. Nucleolin is mainly localized in the nucleus and at the cell surface of cultured tumor cells and endothelial cells of angiogenic vessels in vivo.

Nucleolin, also known as C23, is the major nucleolar protein of exponentially growing eukaryotic cells. It is a ubiquitous conserved multi-domain phosphoprotein found in all vertebrate species. Nucleolin is a multifunctional protein involved in most steps of ribosome biogenesis. It has been implicated in chromatin structure, rDNA transcription, rRNA maturation, ribosome assembly, cytokinesis, nucleogenesis, cell proliferation and growth, and nucleo-cytoplasmic transport.

Rabbit polyclonal anti-Nucleolin antibody recognizes human, mouse, and rat nucleolin. Detection of the nucleolin band by immunoblotting is specifically inhibited with the immunizing peptide.

Immunogen

synthetic peptide corresponding to amino acid residues 2-17 of human nucleolin with C-terminal added cysteine, conjugated to KLH. The corresponding sequence differs by 4 amino acids in rat and mouse nucleolin and by 1 amino acid in the rat homolog protein NRP (nucleolin related protein).

Application

Applications in which this antibody has been used successfully, and the associated peer-reviewed papers, are given below.
Western Blotting (1 paper)

Rabbit polyclonal anti-Nucleolin antibody is used to tag nucleolin for detection and quantitation by immunocytochemical and immunohistochemical (IHC) techniques such as immunoblotting (105 kDa), immunoprecipitation, and immunofluorescence. It is used as a probe to determine the presence and roles of nucleolin in ribosome maturation, cytokinesis, nucleogenesis, cell proliferation and growth, and nucleo-cytoplasmic transport in exponentially growing eukaryotic cells.

Physical form

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

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.

Biochem/physiol Actions

Nucleolin also exhibits auto degradation, DNA and RNA helicase activities, and DNA-dependent ATPase activity. Nucleolin activities may be regulated by proteolysis, methylation, ADPribosylation and phosphorylation by several kinases, including casein kinase II (CK2), p34cdc2 and protein kinase C. Nucleolin might function as a cell surface receptor for lipoproteins, viruses, extracellular matrix, growth factors and other molecules. It interacts with telomerase, which is critical for the nucleolar localization of telomerase.

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
nwg

Documents

Certificate of Analysis

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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

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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N6288 Anti-Nucleolin−Atto 488 antibody produced in rabbit, 1.5-3.0 mg/mL, affinity isolated antibody, buffered aqueous solution

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