Anti-SARS-CoV-1/2 S Protein Antibody, clone 2B3E5 ZooMAb® Mouse Monoclonal
SIGMA/ZMS1076 - recombinant, expressed in HEK 293 cells
Synonym: Coronavirus S protein; Cov-2 S protein; E2; Peplomer protein; S glycoprotein; SARS-CoV 1/2 Spike glycoprotein
Product Type: Chemical
Western Blotting Enhanced Validation-Recombinant Antibody Technology SARS-COV-2 ECD and RBD recombinant protein fragments were probed with Cat. No. ZMS1076, Anti-SARS-CoV-1/2 S Protein, clone 2B3E5 ZooMAb® Mouse Monoclonal (1:1,000 dilution). Proteins were visualized using a Goat Anti-Mouse IgG2a secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates SARS-CoV-1/2 S Protein (~160 kDa).
Western Blotting Enhanced Validation-Recombinant Antibody Technology Lysates from Vero E6 cells infected with SARS-CoV-2 were probed with Cat. No. ZMS1076, Anti-SARS-CoV-1/2 S Protein, clone 2B3E5 ZooMAb® Mouse Monoclonal (1 μg/mL). Proteins were visualized using a Anti-Mouse IgG2a secondary antibody conjugated to HRP and a chemiluminescence detection system (Courtesy of Jeff Johnson Laboratory, Icahn School of Medicine at Mount Sinai (ISMMS).
Affinity Binding Assay Enhanced Validation-Recombinant Antibody Technology 100 mg/mL of recombinant ECD of spike protein (Run 1) or non-specific nucleoprotein (Run 2), or recombinant RBD of spike protein (Run 3) were analyzed by affinity binding assay using Anti-SARS-CoV-1/2 S Protein, clone 2B3 ZooMAb® Mouse Monoclonal. The binding of this antibody to ECD of spike protein displayed a KD value of 3.8 x 10-7, and its binding to RBD of spike protein showed a KD of 7.1 x 10-8, and binding to non-specific nucleoprotein displayed a KD of 4.8 x 10-5.
ELISA Enhanced Validation-Recombinant Antibody Technology Plates were coated with 2 mg/mL of S1 spike protein-mouse Fc fusion recombinant protein. Dilutions of antibodies were subsequently added to compare binding of anti-SARS-CoV-1/2 S protein clone 2B3E5 versus humanized anti-SARS-CoV-1/2 S protein clone hu2B3 antibodies to the recombinant protein as well as positive and negative controls. Detection was completed using Goat Anti-Mouse kappa chain HRP or Goat Anti-human IgG HRP. Both clones 2B3E5 and hu2B3 showed similar binding to the S1 portion of the spike protein (Courtesy of Dr. Thomas Moran, Center for Therapeutic Antibody Development).
Flow Cytometry Enhanced Validation-Recombinant Antibody Technology Staining of one million HEK293 cells transfected with two different expression plasmids for SARS-CoV-2 spike protein was performed using 1 μg/mL of Cat. No. ZMS1076, Anti-SARS-CoV-1/2 S Protein, clone 2B3E5 ZooMAb® Mouse Monoclonal (dotted and dashed lines) or the equivalent amount of a 1 μg/mL of Mouse IgG2a isotype control (dashed lines), followed by a PE-conjugated Anti-Mouse IgG2a secondary antibody (Courtesy of Dr. Thomas Moran, Center for Therapeutic Antibody Development).
Flow Cytometry Enhanced Validation-Recombinant Antibody Technology Two different expression plasmids coding for the SARS-CoV-2 spike protein were used to transfect HEK293 cells and 48 hours later stained with the anti-SARS-CoV-1/2 S protein clone 2B3E5 antibody (purified and supernatant tested). In addition, two isotype control antibodies (M2E10, Py102) were also tested as was a second monoclonal Ab specific for the SARS CoV-1 spike (22C7C4). The 2B3 binds to the spike expressed on the cell surface efficiently. The 22C7C4 binds only to the SARs spike (DNS) (Courtesy of Dr. Thomas Moran, Center for Therapeutic Antibody Development).
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | mouse (recombinant) |
| clone | 2B3E5, recombinant monoclonal |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG2aκ |
| mol wt | calculated mol wt 139.13 kDa (SARS-CoV) |
| calculated mol wt 141.18 kDa (SARS-CoV-2) | |
| observed mol wt ~230 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | SARS virus |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| ELISA: suitable | |
| flow cytometry: 1 μg/mL | |
| western blot: 1:5,000 | |
| UniProt accession no. | P0DTC2  |
| P59594 |
| Application: | Affinity Binding Assay: This antibody bound recombinant extracellular domain of Spike protein with a KD of 3.8 x 10-7 and with the recombinant receptor binding domain with a KD of 4.4 x 10-7. Flow Cytometry Analysis: 1 μg/mL from a representative lot detected SARS-CoV-1/2 S protein in Transfected 293 cells (Courtesy of Dr Thomas Moran, Center for Therapeutic Antibody Development). Western Blotting Analysis: 1 μg/mL from a representative lot detected SARS-CoV-1/2 S Protein in Vero E6 cells +/- SARS-Cov-2 infection (Courtesy of Jeff Johnson Laboratory, Icahn School of Medicine at Mount Sinai (ISMMS). ELISA Analysis: A multiple Dilution from a representative lot detected SARS-CoV-1/2 S Protein in S1 fusion protein (Courtesy of Dr Thomas Moran, Center for Therapeutic Antibody Development). Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-SARS-CoV-1/2 S Protein, clone 2B3E5 ZooMAb, Cat. No. ZMS1076, is a recombinant Mouse monoclonal antibody that detects SARS-CoV-1/2 Spike (S) protein and is tested for use in Affinity Binding Assay, ELISA, Flow Cytometry, and Western Blotting. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. Data presented is the available current product information and provided as-is. These products have not been tested or verified in any additional applications, sample types, including any clinical use. Experimental conditions must be empirically derived by the user. Our Antibody Guarantee only covers tested applications stated herein and conditions presented in our product information and is not extended to publications. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | Recombinant fragment corresponding to full-length extracellular domain from SARS-CoV virus spike protein. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified mouse monoclonal antibody IgG2a in buffer containing PBS with 5% Trehalose without preservatives |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 2B3E5 is a ZooMAb mouse recombinant antibody that specifically detects SARS-CoV-1/2 Spike protein. It targets an epitope within the extracellular domain. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | SARS-CoV-1/2 Spike glycoprotein (UniProt: P59594/P0DTC2; also known as S glycoprotein, E2, Peplomer protein) is encoded by the S gene (Gene ID: 1489668/43740568) in SARS-CoV-1/2 virus. The SARS-CoV-2 is a positive-strand RNA virus that causes severe respiratory syndrome in human. The mature SARS-CoV-2 contains 4 structural proteins: Envelope (E), Membrane (M), Nucleocapsid (N), and the Spike protein (S). E and M proteins help in viral assembly and N protein is needed for RNA synthesis. The S protein is a single-pass type I, homotrimeric, membrane glycoprotein that is responsible for virus binding and entry into host cell. It is synthesized with a signal peptide (aa 1-12), which is subsequently cleaved off to generate the mature protein that contains an extracellular domain (aa 13-1213), a transmembrane domain (aa 1214-1234), and a cytoplasmic domain 1235-1273). The S protein is further processed into S1 (aa 13-685) and S2 (aa 686-1273) subunits by host cell furin. The presence of a furin polybasic cleavage site in S protein from SARS-CoV-2 sets it apart from S protein in SARS-CoV that possesses a monobasic S1/S2 cleavage site. The S1 subunit has the receptor binding domain (RBD; aa 319-541) that mediates entry of SARS-CoV-2 into sensitive cells through the peptidase domain of host Angiotensin-converting enzyme 2 (ACE2) with high affinity (KD = 15 nM). The S2 protein, which is reported to be well conserved, is responsible for membrane fusion. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |