| Description | This product is a human monoclonal antibody that reacts with B5 Env. The antibody displayed high binding affinities to B5 (Kd of 0.2 nM). The mAb inhibited the spread of vaccinia virus as well as variola virus in vitro, protected mice from subsequent intranasal challenge with virulent vaccinia virus, and provided significantly greater protection than that afforded by a previously isolated rat anti-B5 mAb (19C2) or by vaccinia immune globulin. The mAb bound to a conformational epitope between amino acids 20 and 130 of B5. |
| Clonality | Monoclonal |
| Host Species | Human |
| Target Species | Vaccinia Virus (VACV) |
| Epitope | The epitope is located in the amino acids region 20-130. |
| Affinity | 0.2 nM |
| Isotype | IgG1 |
| Expression Species | HEK293F or CHO |
| Conjugation | None |
| Purity | >95%, determined by SDS-PAGE and SEC-HPLC |
| Endotoxin | <1 EU/mg |
| Purification | Protein A affinity purified |
| Sterility | 0.2 μM filtered |
| Formulation | PBS, pH 7.4 |
| Preservation | No preservatives |
| Stabilizer | No stabilizers |
| Storage | Store at 4°C within a week. For longer storage, aliquot and store at -20°C. |
| Application | ELISA; WB; Neut; FuncS |
| Application Notes | In vitro neutralizing activity BS-C-1 cells were infected with about 50 pfu of VACV strain IHD-J. After 2 hours at 37°C, the monolayer was washed, and fresh medium containing indicated amounts of chimpanzee anti-B5 8AH8AL was added. After 48 hours, the monolayers were stained with crystal violet. For the smallpox assay (b), monolayers of BS-C-40 cells in six-well cell culture plates were infected with the Solaimen strain of variola virus at 50 pfu per well in RPMI medium containing 2% FBS. After 1 hour, the medium was aspirated; cells were washed twice and overlaid with RPMI medium containing 25, 2.5, or 0 ug of anti-B5 IgG. The plates were then incubated in a CO2 incubator for 4 days at 35.5°C. Cells were fixed and reacted with polyclonal rabbit anti-variola virus antibody. After incubation with goat anti-rabbit-HRP conjugate, comets were visualized by addition of TrueBlue peroxidase substrate. |
| ELISA | Enzyme-Linked Immunosorbent Assay Protocol |
| WB | Western Blot Protocol |
| FC | Flow Cytometry Protocol |
| Target | B5 Env |
| Alternative Name | B5 envelope glycoprotein; B5; VACV; Vaccinia virus |
| Research Area | Infectious Disease |
