Revision 3

#64509

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:
ELISA

MW (kDa):
124

Source/Isotype:
E. coli

UniProt ID:
#J7RUA5

Description

Human Cas9 (S. aureus) (6xHis-Tag) Recombinant Protein Quantitative Control is derived from full-length Cas9 (S. aureus) with a nuclear localization signal (NLS) and 6xHis-Tag on the carboxy-terminal end. The protein can be used with FastScan™ Cas9 (S. aureus) ELISA Kit #86538 and Cas9 (S. aureus) Matched Antibody Pair #50030 as a quantitative control. Suitable for use in other Cas9 (S. aureus) assays when a control is needed.

Directions for Use

Prior to use, thaw the recombinant protein and briefly spin down to collect all liquid at the bottom of the tube. Dilute according to the following instructions depending on the ELISA kit type. An example sensitivity curve is shown in Figure 2.

PathScan® ELISA Kits:
Dilute with ELISA Sample Diluent to achieve a 1X protein concentration.

PathScan® RP ELISA Kits:
Dilute with 1X Cell Lysis Buffer #9803 to a 2X protein concentration to achieve a final 1X protein concentration upon addition of the detection antibody.

FastScan™ ELISA Kits:
Dilute with 1X FastScan™ ELISA Cell Extraction Buffer to a 2X protein concentration to achieve a final 1X protein concentration upon addition of the antibody cocktail.

Formulation

Expression Host: E. coli

Supplied in 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM DTT, 300 mM NaCl, and 50% (v/v) glycerol.

Storage

Store at -20°C.

Purity

Assessed to be >95% by densitometry using SDS-PAGE analysis and Coomassie Blue detection.

Background

The CRISPR associated protein 9 (Cas9) is an RNA-guided DNA nuclease and part of the CRISPR antiviral immunity system that provides adaptive immunity against extra chromosomal genetic material (1). The CRISPR antiviral mechanism of action involves three steps: (i), acquisition of foreign DNA by host bacterium; (ii), synthesis and maturation of CRISPR RNA (crRNA), followed by the formation of RNA-Cas nuclease protein complexes; and (iii), target interference through recognition of foreign DNA by the complex and its cleavage by Cas nuclease activity (2). The type II CRISPR/Cas antiviral immunity system provides a powerful tool for precise genome editing and has potential for specific gene regulation and therapeutic applications (3). The Cas9 protein and a guide RNA consisting of a fusion between a crRNA and a trans-activating crRNA (tracrRNA) must be introduced or expressed in a cell. A 20-nucleotide sequence at the 5' end of the guide RNA directs Cas9 to a specific DNA target site. As a result, Cas9 can be "programmed" to cut various DNA sites both in vitro and in cells and organisms. CRISPR/Cas9 genome editing tools have been used in many organisms, including mouse and human cells (4,5). Research studies demonstrate that CRISPR can be used to generate mutant alleles or reporter genes in rodents and primate embryonic stem cells (6-8).

Cas9 (S. aureus) is a Cas9 ortholog that is smaller, but as efficient, as the more commonly used Cas9 ortholog, Cas9 (S. pyogenes) (9).

Applications Key

ELISA: ELISA

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 3

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Figure 1. The purity of Cas9 (S. aureus) (6xHis-Tag) Recombinant Protein Quantitative Control is shown by SDS-PAGE staining with Coomassie Blue. Cas9 (S. aureus) (6xHis-Tag) has a predicted molecular weight of 124 kDa.

ELISA Image 1: Cas9 (<i>S. aureus</i>) (6xHis-Tag) Recombinant Protein Quantitative Control
Figure 2. Titration curve demonstrating the relationship between protein concentration of Cas9 (S. aureus) (6xHis-Tag) Recombinant Protein Quantitative Control #64509 and Cas9 (S. pyogenes) (6xHis-Tag) Recombinant Protein Quantitative Control #47140 and the absorbance at 450 nm using the FastScan™ Cas9 (S. aureus) ELISA Kit #86538.
ELISA Image 2: Cas9 (<i>S. aureus</i>) (6xHis-Tag) Recombinant Protein Quantitative Control
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.