SARS-CoV 3CL Protease antibody

Antigen: SARS-CoV 3CL Protease

Clonality: Polyclonal

Host: Rabbit

Application: Immunofluorescence (IF), ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN129545

Additional Information

Gene ID: NP_828863

Swiss-Prot: P59641

Immunogen: This protein A purified antibody was prepared from whole rabbit serum produced by repeated immunizationswith a purified recombinant protein corresponding to full length SARS Coronavirus 3CL Protease. Lifesensors Inc.( www.lifesensors.com ) prepared the 3CL Protease as follows: SUMO-3CL protease fusion was expressed in E. coli in LB medium and purified by Ni-NTA resin affinity chromatography (Qiagen). After the fusion was cleaved by the SUMO Protease(LifeSensors), the SUMO tag and protease were subtracted from the 3CL protease using MAC and the 3CL protease wasfinally purified using Anion Exchange Chromatography with the Macro-Prep High Q resin (BioRad) and size exclusionchromatography.

Format: Liquid (sterile filtered)

Description: Generally, viruses haveproteases to process their proteins into active form.Because of its pivotal role in the viral life cycle,proteases are primary targets for the development ofantiviral agents. 3CL protease, a viral cysteineproteinase, plays an important role in co-translationalproteolytic processing of Coronavirus polyproteins.The 3CL protease cleaves as much as 11 sites in thereplicase polyproteins and also releases the keyreplicative functions of polymerase and helicase. 3CLprotease is the only Coronavirus protein for whichstructural information is available. 3CL proteasecomprises three domains, the substrate-binding site isexpected to be located between domains I and II, anddomain III is a globular cluster comprising five helices.3CL protease is a homodimer.

Specificity: This protein A purified antibody is directed against SARS Coronavirus 3CL Protease. The productwas purified from monospecific antiserum by protein A affinity purification. BLAST analysis was used to suggest reactivity withrelated Coronavirus proteins. Cross reactivity with homologues from other sources has not been determined.

Application Details

Application Notes: This protein A purified antibody has been tested for use in ELISA and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 34 kDa in size corresponding toSARS 3CL Protease by western blotting in the appropriate cell lysate or extract. Dilutions: ELISA 1:10,000 - 1:50,000 WESTERN BLOT 1:2,000 - 1:10,000 IF MICROSCOPY User Optimized OTHER APPLICATIONS User Optimized

Concentration: 5.0 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at 4° C prior to restoration. Reconstitute in 0.1 ml of deionized water or equivalent. Diluteonly prior to immediate use. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing andthawing. Expiration date is one (1) year from date of opening.

Research Area: Virology

Restrictions: For Research Use only

Images

Western blot using antibodies-online's Protein A Purified anti-SARS CoV 3CL Protease antibody shows detection of a 34- kDa band corresponding to the protein. Approx. 100 ng of protein was loaded for SDS-PAGE and transferred onto nitrocellulose. The blot was incubated with a 1:5,000 dilution of the antibody at room temperature for 1 h followed by detection using IRDye