Technical Data
W1018-53
West Nile Virus, NS2b/NS3 Protease (WNV)
Description:
NS3 and NS2b are non-structural viral proteins that play an essential role in viral replication and are therefore a potential target for treatment and prevention of West Nile Virus disease. NS3 is a trypsin-like serine protease that requires NS2b as a cofactor.

Hybridoma:
Produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified, E. coli-derived, recombinant West Nile Virus NS3 Protease complex (rwnvNS3/NS2b; aa 1-184 of NS3; Accession # NP_776018, and aa 49-96 of NS2b; Accession # NP_776017).

Applications:
Western Blot, Neutralization, Immunoprecipitation, Direct ELISA

Recommended Dilution:
Western Blot: This antibody can be used at 1-2ug/mL with the appropriate secondary reagents to detect west nile virus NS3/NS2b complex. Using a colorimetric detection system, the detection limit for rwnvNS3/NS2b complex is approximately 5 ng/lane under non-reducing and reducing conditions. Chemiluminescent detection will increase sensitivity by 5 to 50 fold.

Neutralization of west nile virus NS3 bioactivity: This antibody has been used to inhibit/neutralize the enzymatic activity of rwnvNS3/NS2b. The antibody was preincubated with the enzyme at different molar ratios at room temperature for 30 min. The enzyme was then assayed with a peptide substrate, Ac-REKR-AFC. 50% of the proteolytic activity was inhibited by the antibody at approximately 13ug/mL (IC50) under conditions in which the enzyme was present at 10ug/mL and the substrate concentration was 200uM. Considering the molecular masses of the enzyme (27kD) and the antibody (150kD), IC50 was achieved at approximately 1:4 molar ratio of the antibody to the enzyme.

Immunoprecipitation: This antibody was used at a concentration of 25ug/mL to immunoprecipitate rwnvNS3/NS2b from conditioned cell culture medium. The recovered rwnvNS3/NS2b can be detected in Western blots using a polyclonal antibody.

Direct ELISA: This antibody can be used at 0.5-1.0ug/mL with the appropriate secondary reagents to detect west nile virus NS3/NS2b complex. The detection limit for rwnvNS3/NS2b is approximately 1 ng/well.

Optimal dilutions to be determined by the researcher.

Storage and Stability:
Lyophilized powder may be stored at 4C for short-term only. Reconstitute to nominal volume by adding sterile 40-50% glycerol and store at -20C. Reconstituted product is stable for 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
MabIgG38H153Affinity Purified
SizeStorageShippingSourceHost
100ug4C Do Not FreezeBlue IceMouse
Concentration:
~0.1mg/ml
Immunogen:
E. coli-derived rwnvNS3/NS2b
Purity:
Purified by Protein G affinity chromatography.
Form
Supplied as a lyophilized powder in TBS, 5% trehalose. Reconstitution: Reconstitute with sterile TBS. If 1ml of TBS is used, the antibody concentration will be 100ug/ml.
Specificity:
Recognizes West Nile Virus NS2b subunit.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.