SARS-CoV-2 (2019-nCoV) 3CLpro/3C-like Lysate | 3CLpro lysate
SARS-CoV-2 (2019-nCoV) 3CLpro/3C-like protease-His Recombinant Insect Cell Lysate (WB positive control)
Applications
Western Blot
Synonyms
SARS-CoV-2 (2019-nCoV) 3CLpro/3C-like; SARS-CoV-2 (2019-nCoV) 3CLpro/3C-like protease-His Recombinant Insect Cell Lysate (WB positive control); 3CLpro lysate
Host
Baculovirus-Insect Cells
Form/Format
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Applicable Applications for 3CLpro lysate
Western Blot (WB)
Application Notes
Western blot (WB): Use at an assay dependent dilution.
Species
SARS-CoV-2
Sequence Information
A DNA sequence encoding the SARS-CoV-2 (2019-nCoV) 3CL Protease (YP_009725295.1)(Ser3264-Gln3569) was expressed with a N-terminal polyhistidine tagged.
Preparation Method
Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
3. Store the lyophilized cell lysate at 4 degree C. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80 degree C.
2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
3. Store the lyophilized cell lysate at 4 degree C. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80 degree C.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Related Product Information for 3CLpro lysate
Description: This SARS-CoV-2 CoV 3CLpro/3C-like protease overexpression lysate was created in Baculovirus-Insect Cells and intented for use as a Western blot (WB) positive control. Purification of CoV 3CLpro/3C-like protease protein from the overexpression lysate was verified.
Background: Baculovirus-Insect Cell SARS-CoV-2 (2019-nCoV) 3CLpro/3C-like protease-His Recombinant transfected/overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
3C-like protease (3CLpro) is the main protease of Humann Coronavirus. 3C-like protease (3CLpro) is a key enzyme, as it cleaves several sites to produce non-structural proteins that are essential for genome replication and Coronavirus virion production, such as an RNA-dependent RNA polymerase, a helicase, ribonucleases and 3CLpro itself, from two types of polyproteins (pp1a and pp1ab). SARS-CoV 3CLpro exists as a homodimer and each protomer has an active site.
Background: Baculovirus-Insect Cell SARS-CoV-2 (2019-nCoV) 3CLpro/3C-like protease-His Recombinant transfected/overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
3C-like protease (3CLpro) is the main protease of Humann Coronavirus. 3C-like protease (3CLpro) is a key enzyme, as it cleaves several sites to produce non-structural proteins that are essential for genome replication and Coronavirus virion production, such as an RNA-dependent RNA polymerase, a helicase, ribonucleases and 3CLpro itself, from two types of polyproteins (pp1a and pp1ab). SARS-CoV 3CLpro exists as a homodimer and each protomer has an active site.
References
Tomonari Muramatsu,et al.Autoprocessing mechanism of severe acute respiratory syndrome coronavirus 3C-like protease (SARS-CoV 3CLpro) from its polyproteins.FEBS Journal.2013
Ziebuhr J,Molecular biology of severe acute respiratory syndrome coronavirus. Curr Opin Microbiol.2004
Yang H, et al. The crystal structures of severe acute respiratory syndrome virus main protease and its complex with an inhibitor. Proc Natl Acad Sci USA.2003
Ziebuhr J,Molecular biology of severe acute respiratory syndrome coronavirus. Curr Opin Microbiol.2004
Yang H, et al. The crystal structures of severe acute respiratory syndrome virus main protease and its complex with an inhibitor. Proc Natl Acad Sci USA.2003
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Product Notes
The 3CLpro (Catalog #AAA8119738) is a Lysate produced from Baculovirus-Insect Cells and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's SARS-CoV-2 (2019-nCoV) 3CLpro/3C-like can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). Western blot (WB): Use at an assay dependent dilution. Researchers should empirically determine the suitability of the 3CLpro for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "SARS-CoV-2 (2019-nCoV) 3CLpro/3C-like, Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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