ELISA: Antigen (REC31807) was diluted over the range 1000ng/ml to 0.001ng/ml in GIBCO D-PBS and 100µl volumes added to wells in Greiner high binding strip well plate. Zero wells = D-PBS only. The wells were coated overnight in the fridge. Following morning plate washed 2 X 300µl 1% BSA in D-PBS and allowed to soak/block 1hr 20min ambient. Plate washed 1 X 300µl in D-PBS + 0.1% Tween 20, emptied and used. Antibody (MAB12421) diluted in D-PBS + 1% BSA + 0.05% tween 20, diluted to 1µg/ml and 0.01µg/ml and added at 100µl/well. Zero wells = diluent only. Plate incubated shaken 800rpm 2 hours 20min. Plate washed 3 X 300µl in D-PBS + 0.1% Tween 20, emptied and used. Goat anti-Mouse Ig-HRP (Southern Biotech, cat# 1010-005 lot H0416-ZF86C) diluted 5µl in 11ml D-PBS + 1% BSA + 0.05% Tween 20, added 100µl/well, incubated shaken 1 hour. Plate washed 6 X 300µl in D-PBS + 0.1% Tween 20, emptied and used. TMB (Europa MO701A) added at 100µl/well and incubated 15 min shaken. Plate stopped 100µl/well 1M HCl and read OD at 450nm. A second reading was taken at 405nm to allow estimation of the OD values for wells that were out of range at 450nm.
Mouse Anti SARS-CoV-2 Spike (S2) Antibody (0018)
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Mouse anti SARS-CoV-2 Spike (S2) antibody (0018) is a monoclonal antibody that recognizes the SARS-CoV-2 Spike S2 glycoprotein. This antibody has been manufactured for use in Western blot and ELISA immunoassay development. Antibody does not recognize SARS-CoV-2 Spike S1 glycoprotein.
MOUSE ANTI SARS-COV-2 SPIKE (S2) ANTIBODY (0018)
Mouse anti SARS-CoV-2 Spike (S2) antibody (0018) is a monoclonal antibody that recognizes the SARS-CoV-2 Spike S2 glycoprotein, which is the causative agent of COVID-19. This antibody has been manufactured for use in Western blot and ELISA immunoassay development. Antibody does not recognize SARS-CoV-2 Spike S1 glycoprotein.
PRODUCT DETAILS – MOUSE ANTI SARS-COV-2 SPIKE (S2) ANTIBODY (0018)
- Antibody recognizes SARS-CoV-2 spike S2 glycoprotein.
- Antibody does not recognize SARS-CoV-2 Spike S1 glycoprotein.
- Isotype – Mouse IgG1
- Immunogen was recombinant fragment of glycoprotein precursor expressed in E. coli.
- Protein A or G purified from hybridoma culture supernatant.
- Suitable for use in Western blot and ELISA.
SARS (severe acute respiratory syndrome) is caused by a human coronavirus. Human coronaviruses are the major cause of upper respiratory tract illness, such as the common cold, in humans. Coronaviruses are positive-stranded RNA viruses, featuring the largest viral RNA genomes known to date (27-31 kb). The complete sequence of the SARS coronavirus contains 25 open reading frames. The coronavirus genome encodes four structural proteins: the spike (S) protein, nucleocapsid (N) protein, membrane (M) protein, and the envelope (E) protein.
The spike protein is encoded by the S gene and is a major target of the cellular immune response to coronaviruses and plays an important role in the initial stages of infection. The glycosylated spike protein (as well as the nucleocapsid protein) can be detected in infected cell culture supernatants with antisera from SARS patients (Krokhin et al., 2003). The spike (S) glycoprotein on the coronavirus envelope mediates the attachment of the virus to the cell surface receptors and induces the fusion of the viral and cellular membranes. It is responsible for host cell attachment, receptor binding, and for mediating host cell membrane and viral membrane fusion during infection. Spike is synthesized as a precursor single polypeptide chain of ∼1300 amino acids and then cleaved by host furin-like proteases into an amino (N)-terminal S1 subunit and a carboxyl (C)-terminal S2 subunit. Spike attaches the virion to the cell membrane by interacting with human ACE2 and CLEC4M/DC-SIGNR, initiating the infection. Binding to the receptor and internalization of the virus into the endosomes of the host cell probably induces conformational changes in the S glycoprotein. Proteolysis by cathepsin CTSL may unmask the fusion peptide of S2 and activate membrane fusion within endosomes.Therefore, the S1 subunit, especially its receptor-binding domain (RBD) is critical in determining cell tropism, host range and zoonotic transmission of coronaviruses. Spike protein S2 mediates fusion of the virion and cellular membranes by acting as a class I viral fusion protein. Spike protein S2′ acts as a viral fusion peptide which is unmasked following S2 cleavage occurring upon virus endocytosis (Gui et al., 2017).
- Gui M, Song W, Zhou H, et al. Cryo-electron microscopy structures of the SARS-CoV spike glycoprotein reveal a prerequisite conformational state for receptor binding. Cell Res. 2017;27(1):119–129.
- Krokhin et al. (2003). Mass spectrometric characterization of proteins from the SARS virus: a preliminary report. Mol Cell Proteomics. 2(5):346–356.