Reassessment of CXCR4 Chemokine Receptor Expression in Human Normal and Neoplastic Tissues Using the Novel Rabbit Monoclonal Antibody UMB-2 英文参考文献.docVIP
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Reassessment of CXCR4 Chemokine Receptor Expression in Human Normal and Neoplastic Tissues Using the Novel Rabbit Monoclonal Antibody UMB-2 英文参考文献
ReassessmentofCXCR4ChemokineReceptorExpression
inHumanNormalandNeoplasticTissuesUsingthe
NovelRabbitMonoclonalAntibodyUMB-2
ThomasFischer1,FalkoNagel1,2,StefanJacobs1,RalfStumm3,StefanSchulz1,2*
1Department of Pharmacology, Julius-Maximilians-University, Wu¨rzburg, Germany, 2Department of Pharmacology and Toxicology, Friedrich-Schiller-University, Jena,
Germany,3DepartmentofPharmacologyandToxicology,Otto-von-Guericke-University,Magdeburg,Germany
Abstract
Background:TheCXCR4chemokinereceptorregulatesmigrationandhomingofcancercellstospecificmetastaticsites.
Determination of the CXCR4 receptor status will provide predictive information for disease prognosis and possible
therapeuticintervention.However,previousattemptstolocalizeCXCR4usingpoorlycharacterizedmousemonoclonalor
rabbitpolyclonalantibodieshaveproducedpredominantnuclearandoccasionalcytoplasmicstainingbutdidnotresultin
theidentificationofbonafidecellsurfacereceptors.
Methodology/Principal Findings: In the present study, we extensively characterized the novel rabbit monoclonal anti-
CXCR4 antibody (clone UMB-2) using transfected cells and tissues from CXCR4-deficient mice. Specificity of UMB-2 was
demonstrated by cell surface staining of CXCR4-transfected cells; translocation of CXCR4 immunostaining after agonist
exposure; detection of a broad band migrating at M 38,000–43,000 in Western blots of homogenates from CXCR4-
r
expressingcells;selectivedetectionofthereceptorintissuesfromCXCR4+/+butnotfromCXCR42/2mice;andabolition
of tissue immunostaining by preadsorption of UMB-2 with its immunizing peptide. In formalin-fixed, paraffin-embedded
humantumortissues,UMB-2yieldedhighlyeffectiveplasmamembranestainingofasubpopulationoftumorcells,which
wereoftenheterogeneouslydistributedthroughoutthetumor.Acomparativeanalysisofthemousemonoclonalantibody
12G5andotherfrequentlyusedcommerciallyavailableantibodiesrevealedthatnoneofthesewasabletodetectCXCR4
underotherwiseidenticalconditions.
Conclusions/Significance:Thus,therabb
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