United Kingdom (UK)
Recombinant monoclonal antibody to MBP. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the synthetic construct sAb-70.
UniProt Accession Number of Target Protein: P0AEY0; P0AEX9
Alternative Name(s) of Target: malE; MMBP; Maltose-binding periplasmic protein; Maltodextrin-binding protein
Immunogen: This conformationally selective synthetic antibody was generated by customized phage display selections that stabilize MBP in different conformational states.
Specificity: This antibody is a conformation specific antibody and binds the maltose binding protein in its open conformation. It is reported that sAB-7O binds a large endosteric site on MBP using its heavy-chain CDR-H1, -H2, and -H3 and light chain CDR-L1 and -L3 residues.
Application Notes: This antibody binds MBP with KD of 1 nM in the absence of maltose and 152 nM in 1 µM maltose and does not bind in 1 mM maltose. This antibody binds to the maltose binding pocket of MBP with sufficient affinity to effectively outcompete maltose binding and driving the equilibrium to the open form. Therefore, in presence of excess sAB-7O there was no appreciable observation of maltose binding. Competitive ELISA was used to determine that the open (sAB-7O) and closed conformation (sAB-11M) specific antibodies compete for binding site of MBP (PMID: 29321208).
Antibody first published in:
Mukherjee et al. Engineered synthetic antibodies as probes to quantify the energetic contributions of ligand binding to conformational changes in proteins. J Biol Chem. (2018); 293(8); 2815-2828. PMID:29321208
Note on publication:
Describes the conformational structure and characterization of this antibody.
ELISA using sAb-70 (Ab02007) and Multi Tag protein 2. Binding curves of the mouse IgG1 chimeric version of the Anti-MBP antibody sAb-70 (Ab02007-1.1; grey line) and isotype control (Ab00102-1.1, anti-fluorescein; orange line) to an ELISA plate coated with Multi Tag protein 2 (Pr00565, Absolute Antibody) at a concentration of 5 ug/ml. Ab02007 antibody was titrated on a 3-fold serial dilution starting at 10,000 ng/mL. Anti-mouse IgG was used as secondary antibody for detection at a dilution of 1:4,000.
Immunofluorescence staining of HEK293 cells with Anti-MBP (Ab02007) sAb-70 Immunofluorescence analysis of paraformaldehyde fixed HEK293 cells transfected with MBP expressing plasmid (A) and non-transfected HEK293 cells (B), permeabilized with 0.15% Triton stained with the chimeric rabbit IgG version of sAb-70 (Ab02007-23.0) (1:200 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane and cytoplasmic staining. The nuclear stain is DAPI (blue).