BD Biosciences
Purified Mouse Anti-human HLA-A2, clone BB7.2
551230
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Since first being reported in 1978, monoclonal antibodies with specificity for HLA class I molecules have been invaluable tools for basic and clinical research in human immunology. BB7.2 is an example of a polymorphic antibody that recognizes epitopes carried by subsets of HLA class I variants. BB7.2 is specific for HLA class I allotypes designated HLA-A2 and the allotype HLA-A*69, a variant that is a recombinant of HLA-A*02 and HLA-A*68. The epitope recognized by BB7.2 lies in the alpha-2 domain of the HLA class I molecule, with the residue at position 107 critical for recognition of the antibody.
The monoclonal antibody BB7.2 can be used to stain cells transfected with subsets of HLA-A, primary cells or beads coated with subsets of HLA-A. We have used binding to single antigen beads in our lab to better define the epitope recognized by this antibody.
Single antigen HLA class I beads
50ul of 1ug/ml BB7.2 antibody stock is incubated with single antigen HLA class I beads for 60 mins at 4°C, washed and then stained with an appropriate secondary antibody that recognizes murine IgG2 Fc.
N/A
Goat anti-mouse IgG2 conjugated with PE fluorophore (Santa-Cruz Biotechnology, cat# sc-3738) incubated for 60min, washed and then analyzed on Luminex 100.
Analyzed on a Luminex 100 reader that determines the unique identification of a specific bead and the fluorescence of the secondary antibody (conjugated to PE).
BB7.2 anti-human HLA-A2 binds specifically to all HLA-A*02 allotypes tested in the single antigen panel and also to HLA-A*69. When used at a higher concentration, for example 50ug/ml, some cross reactivity is seen with HLA-A*24 and HLA-A*68, two allotypes that have some sequence similarity to HLA-A*02 and HLA-A*69. At 1ug/ml, there is very little background binding indicating that this antibody is highly purified and specific.
W6/32 is an antibody that recognizes all HLA class I allotypes (monomorphic) and can be included in each experiment to normalize for the antigen density on each individual bead. Results are then presented as relative fluorescence ratios. Given that the epitope of this antibody falls exclusively within the alpha-2 domain of the HLA class I molecule, it is unlikely that the identity of the peptide plays a significant role in recognition by the antibody.
Highly specific monomorphic anti HLA-A2 and A*69 antibody.
Some cross-reactivity noted at higher concentrations.
A well-characterized and specific HLA class I antibody.