21-S2RBD-RCg-F/B1

SARS-CoV-2 Biotinylated Spike glycoprotein Receptor Binding Domain from mammalian CHO cells, frozen

Description: Biotinylated SARS-CoV-2 Spike glycoprotein Receptor Binding Domain (SCoV2-RBD-Biotin) expressed as secreted protein in mammalian (hamster) CHO cells. The construct contains amino acids 319-529 of the Spike glycoprotein provided in UniProtKB sequence accession number: P0DTC2 (SPIKE_SARS2). Whole expression construct includes SCoV2-RBD (aa 319 – 529 of Spike glycoprotein) followed by introduced protease cleavage site and C-terminal His6-tag. SCoV2-RBD protein was purified from culture medium under native (non-denaturing) conditions using convenient chromatographic techniques. Then it was biotinylated by chemical labeling via amines using validated method, buffer changed to PBS, aliquoted and frozen for storage.

Form: Frozen liquid in PBS (pH 7.4).

Storage: The product is shipped with dry ice (shipment cost to Europe: 160 €). Store at -80°C for the long term. After thawing may be stored for short-term at 4°C (stable at least 2 weeks at this temperature). Avoid repeated freeze/thaw cycles.

Concentration: 0.5 mg/ml.

Purity: >97 % as determined by SDS-PAGE.

Image: SDS-PAGE showing SCoV2-RBD-Biotin product 21-S2RBD-RCg-F/B1 (in lane B1) at approximately 31-34 kDa (3 μg/lane). Lane F shows sample of non-labeled SCoV2-RBD product 20-S2RBD-RCg-F used for biotinylating (3 μg/lane), whereas lane M – molecular weight marker (ThermoFisher, Cat. No. #26614).

Applications (tested): ELISA, other immunoassays, SDS-PAGE.

Labeling method and analysis of Biotin incorporation: Labeling of the protein with Biotin was performed by validated method using commercially available Biotinylation Kit. The HABA Assay was used for measuring the level of Biotin incorporation. The biotin to protein ratio in SCoV2-RBD-Biotin product 21-S2RBD-RCg-F/B1 is between 1.5 and 2 (Lot dependent) as determined by the HABA assay.

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