United Kingdom (UK)
Recombinant monoclonal antibody to CCR5. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from phage display ST6.
UniProt Accession Number of Target Protein: P51681
Alternative Name(s) of Target: CD195; C-C chemokine receptor type 5; C-C CKR-5; CC-CKR-5; CCR-5; CHEMR13; HIV-1 fusion coreceptor; B8LFP0
Immunogen: This antibody was isolated from a phage displayed antibody fragment library. The library was generated from a New Zealand White rabbit immunised with purified CCR5-N-GST protein, and four rounds of panning were completed against immobilised CCR5-N-MBP antigen.
Specificity: This antibody is specific for the YTSE region of the N-terminal extracellular domain of human CCR5.It also binds to rhesus CCR5.
Application Notes: When expressed intracellularly, this antibody is able to inhibit the surface expression of human CCR5, conferring T-lymphocytes with immunity to CCR5-dependent HIV-1 infection (Steinberger et al, 2000). Humanised ST6 (ST6/34) also has the same depleting effect (Steinberger et al, 2000). The high affinity of this antibody for CCR5-N-MBP has been confirmed in surface plasmon resonance analysis (Steinberg et al, 2000). This antibody has been used to stain CCR5-expressing cells in flow cytometry, with no staining of control cells observed (Steinberger et al, 2000).
Antibody first published in:
Steinberger et al. Generation and Characterization of a Recombinant Human CCR5-specific Antibody: A PHAGE DISPLAY APPROACH FOR RABBIT ANTIBODY HUMANIZATION J Biol Chem. 2000 Nov 17;275(46):36073-8. PMID:10969070
Note on publication:
Describes the generation of this antibody by phage display, and its use in FC and ELISA.
Flow cytometry using the Anti-CCR5 antibody ST6 (Ab01124). Paraformaldehyde fixed U937 cells were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of ST6 (Ab01124-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.
Western Blot using anti-CCR5 antibody ST6 (Ab01124). K562 cell lysates (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of ST6 (Ab01124-23.0) at 0.1 µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunofluorescence staining of fixed U937 cells with anti-CCR5 antibody ST6 (Ab01124). Immunofluorescence analysis of paraformaldehyde fixed U937 cells on Shi-fix™ cover-slips stained with the chimeric rabbit IgG version of ST6 (Ab01124-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane and cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab01124-23.0, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.