United Kingdom (UK)
Recombinant monoclonal antibody to alpha-5 TMD. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from phage display Nb25.
UniProt Accession Number of Target Protein: P31644; P28472
Alternative Name(s) of Target: GABRA5; Gamma-aminobutyric acid receptor subunit alpha-5; GABA(A) receptor subunit alpha-5
Immunogen: A llama was immunized six times with recombinant 1D4-tagged human a1b3 GABAAR pentamer. Nanobodies were selected by phage display, and the clones were then screened for binding to the receptor. From these clones Nb25 was selected.
Specificity: Binds to the transmembrane domain of the alpha-5 GABAA receptor. To see the exact epitope see the crystal structure (Miller et al, 2017). Nb25 straddles the neurotransmitter-binding site between adjacent subunits.
Application Notes: The antibody was used to facilitate the crystallisation of the a5 GABAA receptor transmembrane domain where it was shown to form a 1080A2 area interface (approx.). Nb25 was used to increase the soluability and aid packing in the crystal. It has been shown that the binding of Nb25 does not affect the potentiation of the receptor. (Miller et al, 2017)
Antibody first published in:
Miller et al Structural basis for GABAA receptor potentiation by neurosteroids Nat Struct Mol Biol. 2017 Nov;24(11):986-992 PMID:28991263
Note on publication:
Reports crystal structures of a chimeric GABAA receptor in both the apo and pregnanolone-bound states. Discusses the generation of this antibody, and its use in crystallisation of the GABAA receptor.