Cell Signaling Technology

Product Pathways - Tag Antibodies

Cas9 (7A9-3A3) Mouse mAb (HRP Conjugate) #97982

sc-392737  

No. Size Price
97982S 100 µl ( 10 western blots ) ¥3,986.00 现货查询 购买询价
97982 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 All Species Expected, Transfected Only 160 Mouse IgG1

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting,

Specificity / Sensitivity

Cas9 (7A9-3A3) Mouse mAb (HRP Conjugate) recognizes transfected levels of total Cas9 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of Cas9 from Streptococcus pyogene.

Description

This Cell Signaling Technology antibody is conjugated to the carbohydrate groups of horseradish peroxidase (HRP) via its amine groups. The HRP conjugated antibody is expected to exhibit the same species cross-reactivity as the unconjugated Cas9 (7A9-3A3) Mouse mAb #14697.

Western Blotting

Western Blotting

Western blot analysis of Cas9 expression in untreated 293 cells (-) or 293 cells transfected with a NPRL2 CRISPR/Cas9 KO plasmid (+) using Cas9 (7A9-3A3) Mouse mAb (Biotinylated) #20083. 293/Nprl2 -/- cells were kindly provided by Rachel Wolfson, Lynne Chantranupong, and David Sabatini of MIT, Cambridge, MA.

Background

The CRISPR associated protein 9 (Cas9) is an RNA-guided DNA nuclease and part of the Streptococcus pyogenes 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).

  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.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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

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