Cas9 detection

The PrecisionX™ Cas9 Detection Systems from System Bioscience (SBI) enable accurate monitoring of the expression of the Cas9 editing enzyme. Capable of detecting and quantitating Cas9 mRNA transcript expression levels, the PrecisionX™ Cas9 Detection Systems are validated qPCR/RT-PCR primer sets. The Cas9 editing enzyme protein itself can also be tracked using the validated Cas9 polyclonal antibody.

Benefits of using these Cas9 Detection Systems
• Accurately detect and quantitate Cas9 mRNA transcript expression levels with Cas9 RT-PCR Primer Sets
• Track Cas9 editing enzyme protein itself using the validated Cas9 polyclonal antibody

Cas9 RT-PCR Primer Sets
Cambridge Bioscience offers two validated sets of RT-PCR primers from SBI to validate the expression of Cas9 mRNA in cells. The amplicon for primer set 1 is 219bp and amplicon for primer set2 is 122bp. Allowing for detection of Cas9 nuclease, Cas9 Nickase and Cas9 double mutant at messenger level, these primer sets are compatible with Cas9 mRNA expression from any SBI Cas9 SmartNuclease construct and vectors from Dr. Feng Zhang's lab.
 

End products separated on a 1.5 percent agarose gel stained with ethidium bromide for PCR products amplified from cDNA generated from either Cas9 transfected cells or nontransfected cells.  The numerical data below the gel image are qPCR Ct values. The Cas9 transfected cells cDNA had a Ct at about 17 and no signal is detectable in nontransfected negative control cells.

Cas9 antibody
The monoclonal Cas9 antibody (Clone 7A9) is designed to detect wild-type Cas9, Nickase (D10A, H840A) and catalytically dead Cas9 (dCas9) in cells expressing the protein. Shown below are Western blot data of nontransfected (A) and Cas9-transfected (B) HEK293 cell protein lysates. This monoclonal antibody may be used for a wide range of assays for detection of Cas9 in target cells, including the following Western blot analysis, immunofluorescence and immunoprecipitation.

Cas9 monoclonal antibody (Clone 7A9) is an IgG1 mouse monoclonal antibody that is Protein G affinity-purified and provided in a solution of 1X PBS (pH7.4), 0.5 percent BSA, 30 percent Glycerol, and preserved in 0.05 percent Sodium Azide. The Cas9 antibody (catalog #CAS9-AB-2) is provided at volume of 25µl and a concentration of 1µg/µl.

Material available for download
Cas9 monoclonal antibody [Clone 7A9] CoA
Cas9 RT-PCR primer set CoA

Note: product availability depends on country - see product detail page.