Revision 9

#86538

Store at +4C

FastScan Cas9 (S. aureus) ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
All

UniProt ID:
#J7RUA5

CST Logo
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.

Product Includes Product # Quantity Color
FastScan™ ELISA Microwell Strip Plate, 96 Well5325796 tests
Cas9 (S. aureus) Rabbit Capture mAb471651 eaGreen (Lyophilized)
Cas9 (S. aureus) Mouse HRP-linked mAb589831 eaRed (Lyophilized)
FastScan™ ELISA Capture Antibody Diluent160763 mlGreen
FastScan™ ELISA HRP Antibody Diluent281203 ml
TMB Substrate700411 ml
STOP Solution700211 ml
Sealing Tape545031 ea
ELISA Wash Buffer (20X)980125 ml
FastScan™ ELISA Cell Extraction Buffer (5X)6990510 ml
FastScan™ ELISA Cell Extraction Enhancer Solution (50X)252431 ml
FastScan™ ELISA Kit #86538 Positive Control Type 2785741 ea

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

For the “C” and “V” kits, the supplied 96-well strip plate consists of twelve 8-well strips in a support frame. This enables custom plate configurations.

Description

The FastScan™ Cas9 (S. aureus) ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Cas9 (S. aureus). To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with Cas9 (S. aureus) in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of Cas9 (S. aureus).

*Antibodies in kit are custom formulations specific to kit.

IMPORTANT: This FastScan™ ELISA Kit requires 4 washes at Step 6 of the protocol.

Specificity/Sensitivity

The FastScan™ Cas9 (S. aureus) ELISA Kit detects endogenous levels of Cas9 (S. aureus), as shown in Figure 1. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

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).

Trademarks and Patents

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

FastScan™ ELISA is a registered trademark of Cell Signaling Technology, Inc.

U.S. Patents 9,086,407, 9,261,500, and 9,476,874, foreign equivalents, and child patents deriving therefrom.

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 9

Cell Signaling Technology Logo
Figure 1. The FastScan™ Cas9 (S. aureus) ELISA Kit #86538 specifically detects Cas9 protein from S. aureus, but not from S. pyogenes. The relationship between recombinant protein concentration and the absorbance at 450 nm using this ELISA kit is shown in the figure. The recombinant Cas9 proteins used were Cas9 (S. aureus) (6xHis-Tag) Recombinant Protein Quantitative Control #64509 and Cas9 (S. pyogenes) (6xHis-Tag) Recombinant Protein Quantitative Control #47140.
ELISA Image 1: FastScan<sup>™</sup> Cas9 (<i>S. aureus</i>) ELISA Kit
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.