Cell Signaling Technology

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Cas9 (7A9-3A3) Mouse mAb (Alexa Fluor® 488 Conjugate) #34963

No. Size Price
34963S 100 µl ( 50 tests ) ¥3,986.00 现货查询 购买询价
34963 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
F 1:50 All Species Expected, Transfected Only Mouse IgG1
IF-IC 1:100

Species cross-reactivity is determined by western blot.

Applications Key: F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Specificity / Sensitivity

Cas9 (7A9-3A3) Mouse mAb (Alexa Fluor® 488 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 Alexa Fluor® 488 fluorescent dye 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 (7A9-3A3) Mouse mAb #14697.

IF-IC

IF-IC

Confocal immunofluorescent analysis of Cas9 expression in NPRL2 CRISPR/Cas9 KO plasmid transfected (left) and untransfected (right) HEK-293 cells using Cas9 (7A9-3A3) Mouse mAb (Alexa Fluor® 488 Conjugate) (green). Cas9 expressing cells were generated by transient transfection of Cas9 and NPRL2-specific guide sequences. Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). HEK-293/NPRL2 knockout cells were kindly provided by Rachel Wolfson, Lynne Chantranupong, and David Sabatini from MIT, Cambridge, MA.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HEK-293 cells untreated (blue) or transfected with NPRL2 CRISPR/Cas9 KO plasmid (green) using

Cas9 (7A9-3A3) Mouse mAb (Alexa Fluor® 488 Conjugate).

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 cell (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

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

Protocols

Companion Products


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

DRAQ5 is a registered trademark of Biostatus Limited.

DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

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

Alexa Fluor is a registered trademark of Life Technologies Corporation.

The Alexa Fluor dye conjugates in this product are sold under license from Life Technologies Corporation, for research use only excluding use in combination with DNA microarrays and high content screening (HCS).

This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is contingent on the buyer using the purchased product solely in research, including use with HCS or other automated imaging applications but excluding use in combination with DNA microarrays. The buyer must not sell or otherwise transfer this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; (c) manufacturing or quality assurance or quality control, or (d) resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.

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

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