C-C Receptor 5/HIV-1 fusion coreceptor/CD195

Accession # AAB65720; Other nomenclature: Chemokine Receptor Type 5, C-C CKR5,HIV-1 fusion coreceptor; CD195

Chemokine receptors represent a subfamily of ~20 GPCRs that were originally identified by their roles in immune cell trafficking.  Macrophage inflammatory protein-1 alpha (MIP-1 alpha) and RANTES, members of the beta chemokine family of leukocyte chemo-attractants, bind to a common seven-transmembrane-domain human receptor.  Chemokine (Chemo-attractant Cytokines) are small peptides that are potent activators and chemo-attractants for leukocyte subpopulations and other non-hemopoietic cells.  Chemokine receptors (CCR) belong to the superfamily of G protein-coupled receptors (GPCR), which regulate the trafficking and activation of leukocytes, and operate as co-receptors in the entry of HIV-1 and directing the proliferation and migration of immature neurons, glia and their precursors (1). Furthermore, chemokine receptors participate in the etiology and progression of various brain disorders, including AIDS dementia, neuro-inflammatory disease and neuroplasia, making them important potential therapeutic targets in these cases.  Induction of chemokines and infiltration of chemokine receptor-bearing cells has also been shown in a variety of animal models of renal diseases, as well as in human diseases and allograft rejection (2).

Significant polymorphism is noted in CCR5 gene.  Identification of the chemokine receptors CCR5 and CXCR4 as the major coreceptors for HIV-1 entry has greatly assisted our understanding of HIV-1 pathogenesis, transmission, and tropism (3). CCR5 and CXCR4 serve as the two major coreceptors for viruses belonging to HIV-1 subtypes A, B, C, D, E, and group O, whereas other chemokine receptors such as CCR2b and CCR3 play only a minor role in facilitating viral entry into stimulated PBMCs.  The coreceptor usage is determined by the viral phenotype rather than its genotype. Binding of the HIV-1 envelope glycoprotein gp120 to the CCR5 coreceptor reduces constraints on the metastable transmembrane subunit gp41, thereby enabling gp41 refolding, fusion of viral and cellular membranes, and infection (4).   CCR5 is a multi-pass membrane protein, containing 7TMDs with apparent MW of 46kDa (373 aa) expressed on many different blood cells. The CCR5 protein has putative N-glycosylation sites near the extra cellular N-terminal end of the proteins.  The protein has a large 3^rd intra cellular loop which interacts with G-proteins.  The short carboxy terminal is intracellular and has putative post-translational sites.

The CCR5-selective antibodies were generated against conserved but unique sequences from N-terminal region of human CCR5 receptor proteins.  The peptide sequence resides between aa 1-35 aa of human CCR5 and are unique to CCR5 protein.  Antibodies generated against this peptide are specific for CCR5, the CCR5 peptide antibodies are affinity purified over immobilized antigen based chromatography, and the purified immunoglobulins are stabilized in antibody stabilization buffer. The affinity purified antibodies are also conjugated to FITC and biotin for direct applications in IHC and cell sorting experiments.  FabGennix Int. Inc., also provide limited quantities of antigenic blocking peptides for CCR5 antibodies.  Antibodies to CCR5 will label CCR5 protein in humans, chimpanzee, and monkey receptor in different protocols.  The antibodies will not label rat and mouse protein. Limited quantities of antigenic blocking peptide and Western blot positive control for CCR5 antibodies are also available in ready-to-use buffer for SDS-PAGE and western blotting experiments. FabGennix Inc., provides custom antibody services to those who are looking for high affinity low background antibodies in many different hosts. 

Immunogen:            Synthetic peptides taken form near N-terminal region of the human CCR5 protein from within aa region 1-35.  The peptide was post-synthetically modified and conjugated to carrier protein to achieve desired antigenicity.  The CCR5 peptide sequence was conserved in human, gibbon, chimp,  and monkey but not in other species  

Concentration:         CCR5-501 antibodies concentration range form 0.64-0.70 mg/ml in antibody stabilization buffer.               

Applications:           Antibodies CCR5-501AP is ideal for WB, IHC and ELISA applications, other applications have not been tested.  These antibodies do not cross react to other chemokine receptors.  The species cross reactivity for this antibody is not fully examined.  The dilutions for this antibody is for reference only, investigators are expected to determine optimal conditions for specific assay.  WB: > 1:500; IMM & i.p pull-down assays: n.d; IHC: n.d.  N.D does not necessarily means the antibody in not suitable for that application, investigators using this antibody in applications not listed here can ask for a complimentary sample.

Reactivity:            This antibody detects a single apparent MW of 45kDa of CCR5 in PC-CCR5 samples.  The antibody does not cross reacts with other members of the CCR family.    

Protocols:            Standard protocol for various applications (WB, IMM and IHC) of this antibody can be obtained upon request from FabGennix Int. Inc., technical service, however, FabGennix Inc., strongly recommends investigators to optimize conditions for use of this antibody.     

Form/Storage:       The antiserum is supplied in antibody stabilization buffer with 0.02% sodium azide as preservative.  For long-term storage of antibodies, store at -20oC.  FabGennix Inc. does not recommend storage of very dilute antibody solutions unless they are prepared in specially formulated multi use antibody dilution buffer (Cat # DiluOBuffer).  Working solutions of antibodies in DiluOBuffer should be filtered through 0.45u filter after every use for long-term storage.

 

For research use only, not for diagnostic or therapeutic use.