Mouse Monoclonal Antibody to SARS-CoV2 S-Protein ACE2 Binding Domain
In late 2019 a novel infectious disease was discovered in Wuhan, China which was quickly recognized to be caused by a previously unknown RNA coronavirus. The virus was very rapidly isolated, the full RNA sequence determined and put on-line on the 10th of January 2020. The sequence revealed that the virus was most closely related to certain bat coronaviruses and the severe acute respiratory syndrome (SARS) coronavirus. Immediately biotechnology companies and research institutes used the RNA sequence information to generate vaccine candidates. The SARS virus was known to enter and infect human cells by means of the so-called spike or S-protein which binds to the extracellular domain of the angiotensin converting enzyme 2 (ACE2) protein, which is then internalized bringing the virus into the cell. Cryoelectron microscopy and binding studies quickly determined that the S-protein of SARS-CoV2 is structurally similar to to that of the SARS virus and also binds to the ACE2 receptor, albeit with higher affinity than the S-protein of SARS. This focuses attention on the ACE2 binding site on the SARS-CoV2 S-protein and for the complementary region on ACE2 which binds the SARS-CoV2 S-protein. We therefore expressed both these regions in E. coli, our products PROT-R-SARS-CoV2-bd and PROT-R-ACE2-bd and raised antibodies to them.
The MCA-2G1 antibody was made against our recombinant construct comprising amino acids 308-541 in the S-protein sequence in SARS-CoV2 Wuhan-Hu-1, complete genome. The antibody works well on western blots of crude homogenates of HEK293 cells transfected with the SARS-CoV2 binding domain, cleanly producing the appropriate sized band and as expected also binds the full length S-protein. In addition S-protein transfected cells show clean and strong immunofluorescence staining of the expressed protein with this antibody. We are currently determining the exact peptide epitope of this and our other SARS-CoV2 S-protein antibodies and also measuring their kinetic properties. EnCor supplies another mouse monoclonal antibody to the SARS-CoV2 S-protein ACE2 binding domain MCA-5G8 and also a rabbit polyclonal RPCA-SARS-CoV2-bd. Mouse select image above left for larger view.
HGNC name(s) : N/A
Host : Mouse
Clonality : Monoclonal
Reactivity : Human | Horse | Cow | Pig | Chicken | Rat | Mouse
Isotype : IgG
Conjugation : none
Immunogen : Recombinant SARS-CoV2 S-Protein ACE2 binding domain expressed in and purified from E. coli, EnCor product PROT-r-SARS-CoV2-bd
Mass of detected protein : S-Protein 142kDa
Uniprot ID : P0DTC2
RRID # : AB_2861173
Purification : Purified antibody at 1mg/mL in 50% PBS, 50% glycerol plus 5mM NaN3
Storage : Shipped on ice. Store at 4°C for short term, for longer term at -20°C. Avoid freeze / thaw cycles.
Validated applications : WB | IF/ICC
WB: 1:1 000 – 1:3 000. ICC/IF: 1:1000
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5. Wang, D-S et al. The pleckstrin homology domain of human β-I σ-II spectrin is targeted to the plasma membrane in vivo. Biochem. Biophys. Res. Comm. 225:420-6 (1996).
50 ul, 100 ul, 500 ul
|Validated Applications|| |