Alternative names:
SARS-CoV-2 Omicron variant

Data and images for Recombinant SARS-CoV-2 Spike-Prot B.1.1.529/BA.1 (HEK)

Figures

Figure 1

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Cell-based spike protein binding assay.
Schematic overview illustrating how our biotinylated spike proteins can be applied to analyze binding to ACE2-expressing cells.

Figure 1

Cell-based spike protein binding assay.
Schematic overview illustrating how our biotinylated spike proteins can be applied to analyze binding to ACE2-expressing cells.

Figure 2

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Results of the cell-based binding assay using biotinylated Recombinant SARS-CoV-2 Spike-Prot B.1.1.529/BA.1 (HEK).
ACE2-expressing cells (Vero E6 cells) were incubated with biotinylated SARS-CoV-2 spike monomer of the wild-type, Wuhan variant (Recombinant SARS-CoV-2 Spike-Prot (HEK)-Biotin) and the SARS-CoV-2 variant B.1.1.529/BA.1, Omicron (Recombinant SARS-CoV-2 Spike-Prot B.1.1.529/BA.1 (HEK)-Biotin).
Different protein concentrations were used, and ACE2-bound spike protein was subsequently detected by fluorescent staining with streptavidin-PE. Quantification of stained cells at different protein concentrations illustrates that Miltenyi Biotec's recombinant spike monomer is biologically functional and shows a titratable, high-affinity binding to cell surface ACE2.

Figure 2

Results of the cell-based binding assay using biotinylated Recombinant SARS-CoV-2 Spike-Prot B.1.1.529/BA.1 (HEK).
ACE2-expressing cells (Vero E6 cells) were incubated with biotinylated SARS-CoV-2 spike monomer of the wild-type, Wuhan variant (Recombinant SARS-CoV-2 Spike-Prot (HEK)-Biotin) and the SARS-CoV-2 variant B.1.1.529/BA.1, Omicron (Recombinant SARS-CoV-2 Spike-Prot B.1.1.529/BA.1 (HEK)-Biotin).
Different protein concentrations were used, and ACE2-bound spike protein was subsequently detected by fluorescent staining with streptavidin-PE. Quantification of stained cells at different protein concentrations illustrates that Miltenyi Biotec's recombinant spike monomer is biologically functional and shows a titratable, high-affinity binding to cell surface ACE2.

Figure 3

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SDS-PAGE of biotinylated Recombinant SARS-CoV-2 Spike-Prot B.1.1.529/BA.1 (HEK)
under reduced (R) and non-reduced (NR) conditions.

Figure 3

SDS-PAGE of biotinylated Recombinant SARS-CoV-2 Spike-Prot B.1.1.529/BA.1 (HEK)
under reduced (R) and non-reduced (NR) conditions.

Figure 4

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Size-exclusion chromatography (SEC) Recombinant SARS-CoV-2 Spike-Prot B.1.1.529/BA.1 (HEK).
The non-biotinylated protein was analyzed and the peak corresponds to the monomeric protein.

Figure 4

Size-exclusion chromatography (SEC) Recombinant SARS-CoV-2 Spike-Prot B.1.1.529/BA.1 (HEK).
The non-biotinylated protein was analyzed and the peak corresponds to the monomeric protein.

Specifications for Recombinant SARS-CoV-2 Spike-Prot B.1.1.529/BA.1 (HEK)

Overview

The SARS-CoV-2 spike glycoprotein (S) mediates the entry of the virus into target cells via its interaction with the angiotensin-converting enzyme 2 (ACE2) receptor, thereby initiating the infection. It forms a homotrimeric structure on the surface of the SARS-CoV-2 virus and represents the major target for diagnostic and therapeutic agents. The ectodomain of the S glycoprotein comprises the N-terminal S1 subunit, including the receptor binding domain (RBD) and the C-terminal S2 subunit. The ectodomain spans from aa12 to aa1213 followed by a transmembrane helix beginning at aa1214–aa1234. The C-terminus of the native protein is formed by a cytoplasmic domain spanning aa1235–aa1273.
The SARS-CoV-2 mutant strain B.1.1.529, also known as Omicron variant, was first identified during the Covid-19 pandemic Botswana/ South Africa in November 2021. Due to increased transmissibility and substantial immune evasion it spread rapidly around the globe. The B.1.1.529 lineage and its sublineages are the most mutated variants of the SARS-CoV-2 virus known so far. The BA.1 lineage is defined by 30 amino acid substitutions (A67V, T95I, G142D (already known from B.1.617.2-Delta), L212I, G339D, S371L, S373P,S375F, K417N (already known from B.1.351 lineage Beta), N440K, G446S, S477N, T478K (already known from B.1.617.2-Delta), E484A, Q493R, G496S, Q498R, N501Y (already known from B.1.1.7-Alpha, B.1.351-Beta and P.1-Gamma), Y505H, T547K, D614G (already known from B.1), H655Y, P681H (already known from B.1.1.7-Alpha), N679K, N764K, D796Y, N856K, Q954H, N969K, L981F), 6 amino acid deletions (H69del, V70del (already known from B.1.1.7-Alpha), V143del, Y144del, Y145del, N211del) and one insertion (ins214EPE) in the spike protein. Preliminary evidence suggests an increased risk of reinfection with this variant, as compared to other VOCs.
The Recombinant SARS-CoV-2 Spike-Prot B.1.1.529 (HEK) protein covers the ectodomain of the viral surface protein including amino acids V16 to K1211. It was engineered to contain the stabilizing proline substitutions at position K986P and V987P. The native sequence of the Furin cleavage site (RRAR at residues 682–685) was substituted by GSAG to further increase recombinant protein stability.
The protein is extended at its C-terminus with a His-tag and an AviTag™. Recombinant SARS-CoV-2 Spike-Prot B.1.1.529 (HEK)-Biotin is specifically biotinylated at a single site, preserving full functionality of the protein.

Alternative names

SARS-CoV-2 Omicron variant

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