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Cas9 (S. aureus) (E4G3U) Rabbit mAb (PE Conjugate) #97115

Item# Description List Price Web Price Qty
97115S Cas9 (S. aureus) (E4G3U) Rabbit mAb (PE Conjugate) - 100 µl $427.00
$384.30
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Companion Products

Item# Description List Price Web Price Qty
51610S Cas9 (S. aureus) (E4G3U) Rabbit mAb - 100 µl $364.00
$327.60
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Application Dilution Species-Reactivity Sensitivity Isotype
F Transfected Only Rabbit IgG

Applications Key: F=Flow Cytometry

Protocols

Specificity / Sensitivity

Cas9 (S. aureus) (E4G3U) Rabbit mAb (PE Conjugate) recognizes transfected Cas9 (S. Aureus) protein. This antibody does not cross-react with Cas9 (S. pyogenes), AsCpf1 (Strain BV3L6), and FnCpf (Strain U112).

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val905 of S. aureus Cas9 protein.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of 293T cells, mock transfected (blue) or transfected with a construct expressing myc-tagged Cas9 (S. aureus) (green), using Cas9 (S. aureus) Rabbit mAb (PE Conjugate) (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (PE Conjugate) #5742 (dashed line).

Description

This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometric analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Cas9 (S. aureus) (E4G3U) Rabbit mAb #51610.

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).

  1. Horvath, P. and Barrangou, R. (2010) Science 327, 167-70.
  2. Wiedenheft, B. et al. (2012) Nature 482, 331-8.
  3. Singh, P. et al. (2015) Genetics 199, 1-15.
  4. Cong, L. et al. (2013) Science 339, 819-23.
  5. Mali, P. et al. (2013) Science 339, 823-6.
  6. Li, D. et al. (2013) Nat Biotechnol 31, 681-3.
  7. Shen, B. et al. (2013) Cell Res 23, 720-3.
  8. Niu, Y. et al. (2014) Cell 156, 836-43.
  9. Ran, F.A. et al. (2015) Nature 520, 186-91.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


 

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