E protein (DIII) Recombinant Protein

E protein (DIII) recombinant

Cat. Num. AAA505031

• Applications: ELISA
• Synonyms: E protein (DIII); E protein (DIII) recombinant; RECOMBINANT ANTIGEN E from West Nile Virus; E protein (DIII) recombinant protein

• Host: Escherichia Coli
• Form/Format: Dry Powder; 20mM phosphate buffer pH 8, 1M NaCl, 0.1% polyoxyethylene (10) tridecyl ether and 6 M urea
• Concentration: 1.762 mg/ml (varies by lot)
• Sequence Length: 3433

• Applicable Applications for E protein (DIII) recombinant protein: ELISA (EIA)
• Application Notes: Suggested Titer by ELISA: 1:1855, which corresponds to 0.95 ug/ml in plates for IgG, and IgM detection.; ; Possible Applications: WB, DB, Indirect ELISA, positive control in direct ELISA. Where this product has not been tested for use in a particular technique , this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates.
• Recombinant Antigen: West Nile virus antigen E (Beasley et al., 2004).
• Molecular Weight Information: Determined by SDS-PAGE, the protein band is between molecular markers of 25,000-18,400 Da, while relative molecular mass calculated from amino acid sequence is 19,582.7 Da
• Total Quantity Per Aliquot: 1mg
• Protein Concentration Determined Espectrophotometrically: DO280 = 1.01; A 0.1 % (=1 g/l) = 0.585
• Observation Notes: Proteins should be maintained frozen at high concentrations. The dilution to be performed for ELISA assays should be made with a small quantity of protein, the same day of the experiment. In order to defrost the protein, maintain the aliquot at 25 degree C without shaking to avoid aggregation. Prior making test dilutions and after defrost the protein, is recommended to remove possible protein aggregates by centrifuging the stock solution, avoiding alterations in the immobilization of the biomolecule to the solid surface.; ; This protein is highly prone to micro-aggregation in absence of 6 M urea, therefore we strongly recommend using fresh protein dilutions in ELISA assay for the protein coating in the microtiter plates. Avoid using storageprotein solutions at 4ºC in absence of 6 M urea. As the pI of the protein is 7, we recommend using carbonate buffer for ELISA plates coating.
• Batch Composition: his-E: Recombinant antigen with a his-tag in its N-terminus
• Note: It is ok with the absence of precipitation after a freezing and thawing cycle.
• Reconstitution: With approx. 0.5 ml of double-distilled water, a final concentration of approx. 1.762 mg/ml will be obtained. The solubilisation of the cake protein should be developed for 15 min to allow a homogeneous protein solution , considering that part of the cake can be on the glass-walls of the conatiner. Upon reconstitution , leave the solution at least 15 min homogenizing with a mild agitation at 4°C. Avoid vigorous shaking that can cause foaming and protein denaturation. With this reconstitution , the protein will be maintained at pH 8. It is recommended that the users carry out their absorbance determinations to avoid concentration variabilities due to the equipment used, mainly in reproducibility analysis.
• Preparation and Storage: Protein is shipped at room tempeture. Upon arrival , it should be stored at 4°C or -20°C. Once reconstituted , it should be aliquoted in order to avoid repeated freezing and thawing cycles and stored at -20°C to -80°C.

SDS-PAGE

(SDS-PAGE analysis (15%) of 2 ?l of recombinant E. Purity is > 98% as determined by gel electrophoresis.)

Testing Data #2

Testing Data #3

Related Product Information for E protein (DIII) recombinant protein

Description: The structural domain III of the WNV envelope glycoprotein has been prepared as a recombinant antigen fused to a his-tag at the N-terminal. It is produced from the envelope protein of the West Nile virus.
The measurement of the protein concentration has been performed with the theoretical extinction coefficient of the recombinant protein obtained from Gill and vonHippel, 1989. It is recommended that the users carry out their absorbance determinations to avoid equipment variabilities regarding final concentration, mainly in reproducibility analysis.

Product Categories/Family for E protein (DIII) recombinant protein

Viruses

References

Beasley et al., 2004. Use of a recombinant anvelope protein subunit antigen for specific serological diagnosis of West Nile virus infection. The Journal of Clinical Microbiology, 42:2759-2765.
Gill SC, von Hippel PH. Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989 Nov 1;182(2):319-26.

NCBI and Uniprot Product Information

• NCBI GI #: 377814031
• NCBI Official Full Name: polyprotein