Revision 1

#99098

Store at -20C

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W

Reactivity:
Vir

Sensitivity:
Endogenous

MW (kDa):
60

Source/Isotype:
Rabbit

Entrez-Gene Id:
43740578

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity/Sensitivity

SARS-CoV-2 3'-5' Exonuclease Antibody recognizes endogenous levels of total SARS-CoV-2 3'-5' exonuclease protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val125 of SARS-CoV-2 3'-5' exonuclease protein. Antibodies are purified by peptide affinity chromatography.

Background

The cause of the COVID-19 pandemic is a novel and highly pathogenic coronavirus, termed SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2). SARS-CoV-2 is a member of the Coronaviridae family of viruses (1). The genome of SARS-CoV-2 is relatively large and encodes up to 29 open reading frames (ORFs). These include ORF1a and ORF1b (further processed into 16 non-structural proteins), 9 accessory proteins, and 4 canonical structural proteins: spike (S), envelope (E), membrane (M), and nucleocapsid (N) (2).

SARS-CoV-2 3’-5’ exonuclease, or ExoN, is a non-structural protein (Nsp14) translated from ORF1b (2). It is a proofreading enzyme that functions in the replicase holoenzyme to read, excise, and repair errors in the newly transcribed viral RNA (3). RNA viruses generally have a high mutation rate and Coronaviridae, and others among the Nidovirales genus, have the largest and most complex genomes of RNA viruses (4). The 3’-5’ exonuclease is highly conserved between Coronaviruses, suggesting it’s important to the maintenance of such a large genome (5). The 3’-5’ exonuclease also provides a functionality to the RNA capping process, alongside the 2’-ribose methyl transferase (Nsp14), the helicase (Nsp13), and a guanylyl transferase (6,7). It is, however, the exonuclease function that is the key challenge to antiviral drug design with nucleoside analogue approaches (8).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting

Cross-Reactivity Key

Vir: Virus

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

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Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with plasmid encoding Myc/DDK-tagged SARS-CoV-2 3'-5' exonuclease protein (lane 2), using SARS-CoV-2 3'-5' Exonuclease Antibody (upper), DYKDDDDK Tag (D6W5B) Rabbit mAb #14793 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: SARS-CoV-2 3’-5’ Exonuclease Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.