Anti-SARS-CoV-1/2 S Protein Antibody, clone hu2B3E5 ZooMAb® Chimeric Monoclonal
SIGMA/ZHU1076 - 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
10 ng (Lane 1) and 1 ng (Lane 2) of SARS-CoV-2 spike protein recombinant ECD fragment was probed with 1:1,000 dilution of Cat. No. ZHU1076, Anti-SARS-CoV-1/2 S Protein clone hu2B3E5 ZooMAb® Human Monoclonal (10 ng Lane 1 and 1 ng Lane 2). Proteins were visualized using a Goat Anti-Human IgG1 secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates SARS-CoV-1/2 S Protein (~160 kDa).
Enhanced Validation-Recombinant Antibody Technology
10 ng (Lane 1) and 1 ng (Lane 2) of SARS-CoV-2 spike protein recombinant ECD fragment was probed with 1:1,000 dilution of Cat. No. ZHU1076, Anti-SARS-CoV-1/2 S Protein clone hu2B3E5 ZooMAb® Human Monoclonal (10 ng Lane 1 and 1 ng Lane 2). Proteins were visualized using a Goat Anti-Human IgG1 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
10 ng of SARS-CoV-2 spike protein Receptor Binding Domain (RBD) fragment was probed with 1:1,000 dilution of Cat. No. ZHU1076, Anti-SARS-CoV-1/2 S Protein clone hu2B3E5 ZooMAb® Human Monoclonal (10 ng). Proteins were visualized using a Goat Anti-Human IgG1 secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates SARS-CoV-1/2 S Protein (~45 kDa).
Enhanced Validation-Recombinant Antibody Technology
10 ng of SARS-CoV-2 spike protein Receptor Binding Domain (RBD) fragment was probed with 1:1,000 dilution of Cat. No. ZHU1076, Anti-SARS-CoV-1/2 S Protein clone hu2B3E5 ZooMAb® Human Monoclonal (10 ng). Proteins were visualized using a Goat Anti-Human IgG1 secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates SARS-CoV-1/2 S Protein (~45 kDa).
Affinity Binding Assay Enhanced Validation-Recombinant Antibody Technology 100 mg/mL of the extracellular domain (ECD) of the spike protein (Run 1) or negative control Nucleoprotein (Run 2), or the recombinant receptor binding domain (RBD) of spike protein (Run 3) were analyzed by affinity binding assay using Cat. No. ZHU1076, Anti-SARS-CoV-1/2 S Protein, clone hu2B3E5 ZooMAb® Human Monoclonal. The binding of this antibody to ECD of spike protein displayed a KD value of 4.2 x 10-6, and its binding to RBD of spike protein showed a KD of 9.5 x 10-7, whereas its binding to non-specific nucleoprotein displayed a KD of 5.1 x 10-3.
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 hu2B3E5 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 2B3 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 293expi cells were transfected with SARS-CoV-2 spike protein using expression plasmid and then incubated with antibody dilutions as indicated of anti-SARS-CoV-1/2 antibody clones hu2B3E5 and CR3022. Detection of binding was accomplished using Goat Anti-Human IgG FITC (Courtesy of Dr. Thomas Moran, Center for Therapeutic Antibody Development).
Flow Cytometry Enhanced Validation-Recombinant Antibody Technology Staining of one million HEK293 cells transfected to express spike protein was performed using a several dilutions of Cat. No. ZHU1076, Anti-SARS-CoV-1/2 S Protein clone hu2B3E5 ZooMAb® Human Monoclonal (dashed and dotted lines) or the equivalent amount of a Human IgG1 isotype control (solid lines), followed by a PE-conjugated Goat Anti-Human IgG1 secondary antibody (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 | human (recombinant) |
| clone | hu2B3E5, 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 | IgG1 |
| mol wt | calculated mol wt 139.13 kDa (SARS-CoV) |
| calculated mol wt 141.18 kDa (SARS-CoV-2) | |
| observed mol wt ~160 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: suitable | |
| western blot: suitable | |
| UniProt accession no. | P0DTC2  |
| P59594 |
| Application: | Affinity Binding Assay: A representative lot of this antibody bound recombinant extracellular domain (ECD) of Spike protein with a KD of 4.2 x 10-6 and with the recombinant receptor binding domain (RBD) with a KD of 9.5 x 10-7. Flow Cytometry Analysis: A representative lot detected SARS-CoV-1/2 S Protein in 293 cells transfected to express spike protein (Courtesy of Dr Thomas Moran, Center for Therapeutic Antibody Development). ELISA Analysis: A representative lot detected SARS-CoV-1/2 S Protein in ELISA application. (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 hu2B3E5 ZooMAb, Cat. No. ZHU1076, is a chimeric recombinant Hu Monoclonal antibody (hu Fc region) that detects SARS-CoV-1/2 S Protein and is tested 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 human monoclonal antibody IgG1 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 hu2B3E5 is a chimeric ZooMAb monoclonal antibody that specifically detects SARS-CoV-1/2 Spike protein. It targets an epitope within the extracellular domain. ZHU1076 combines variable regions of mouse monoclonal antibody anti-SARS-CoV-1/2 Spike protein (ZMS1076) and constant region of human IgG1. We recommend an anti-human antibody as secondary antibody for detection (Example: AP309P Goat anti-human IgG antibody HRP). |
| 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. This chimeric version was generated using FC region of human IgG. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |