Arg319 - Gln690. Starting from RBD to C-terminal. Amino acid Asp (D614) was mutated to Gly (G614).
Expression System
HEK293 cells
Tag
C-terminal his-tag
Purification
His-tag affinity purification by immobilized metal ion affinity chromatography (IMAC)
Purity
>90%
Purity Determined By
SDS-PAGE under reducing conditions and visualized by Coomassie blue staining
Molecular Weight (kDa)
Recombinant protein product has a calculated molecular mass of ∼42 kDa. Due to the abundant glycosylation, it migrates as approximately ∼55-60 kDa protein band smear in SDS-PAGE under DTT, beta-mercaptoethanol reducing conditions. After deglycosylation under native and denature conditions, the protein presented as one major ∼42 kDa band. See deglycosylation analysis image below.
Format
Liquid
Formulation
Supplied as a 0.2 µm filtered solution in PBS (pH 7.4)
Concentration (lot specific)
Lot specific (see the label on the vial), determined by BCA protein assay
Endotoxin Level
< 0.5 EU per µg of the protein as determined by the LAL method
Estimated Lead Time
1-2 business days
Shipping Type
Blue ice
Storage
-20°C
SDS-PAGE Image
Figure 1. Deglycosylation analysis of purified recombinant proteins. The same amount of purified proteins were untreated (Lane 2) or treated with protein deglycosylation enzymes under native (Lane 3) or reducing (Lane 4) conditions. Deglycosylation treatment resulted in a mobility shift of the protein to produce one reduced band at the expected size (˜42 kDa), thus indicating that the untreated recombinant protein (Lane 2) was glycosylated. Lane 1: protein standard ladder (kDa). Lane 2: untreated protein. Lane 3: treated protein with deglycosylation enzymes under native conditions. Lane 4: treated protein with deglycosylation enzymes under denature conditions. Lane 5: deglycosylation enzymes only without target proteins.
Binding Function
Product binding ability was measured by ELISA. The immobilized recombinant human ACE2 protein (Catalog #230-30165, coated at 0.5 µg/ml, 100 µl/well) was incubated with the serial diluted SARS CoV-2 S1 RBD D614G mutant protein (Catalog #230-30186). The bound mutant protein was detected by mouse anti-S1 RBD monoclonal antibody using ELISA. The calculated EC50 is 303.5 ng/ml.
Stability & Storage
Shipping
Ice packs
Storage/Stability
Upon arrival, the protein may be stored for 2 weeks at 4 °C. For long term storage, it is recommended to store at -20 °C or -80 °C in appropriate aliquots. Avoid repeated freeze-thaw cycles.
Custom Labeling Request
We can custom label this protein with biotin or fluorescent dyes (choose from violet, blue, green or red). Request now.
