Product Pathways - Tag Antibodies
Cas9 (7A9-3A3) Mouse mAb #14697
|14697S||100 µl ( 10 western blots )||￥3,100.00 现货查询||购买询价|
|14697T||20 µl ( 2 western blots )||￥1,200.00 现货查询||购买询价|
|14697||carrier free & custom formulation / quantity||email request|
|W||1:1000||All Species Expected,||Transfected Only||160||Mouse IgG1|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),
Specificity / Sensitivity
Cas9 (7A9-3A3) Mouse mAb 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.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Cas9 (+), using Cas9 (7A9-3A3) Mouse mAb (upper) and β-Actin (13E5) Rabbit mAb #4970 (lower).
Confocal immunofluorescent analysis of Cas9 expresssion in Nprl2-deficient (left) and untreated 293 cells (right) using Cas9 (7A9-3A3) Mouse mAb (green). Nprl2 expression was knocked out in the Nprl2-deficient cells 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). 293/Nprl2 -/- cells were kindly provided by Rachel Wolfson, Lynne Chantranupong, and David Sabatini of MIT, Cambridge, MA.
Immunohistochemical analysis of Cas9 expresssion in Nprl2-deficient 293 (left, positive) and untreated 293 (right, negative) cell pellets using Cas9 (7A9-3A3) Mouse mAb (green). Nprl2 expression was knocked out in the Nprl2-deficient cells by transient transfection of Cas9 and Nprl2-specific guide sequences. 293/Nprl2 -/- cells were kindly provided by Rachel Wolfson, Lynne Chantranupong, and David Sabatini of MIT, Cambridge, MA.
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).
- Horvath, P. and Barrangou, R. (2010) Science 327, 167-70.
- Wiedenheft, B. et al. (2012) Nature 482, 331-8.
- Singh, P. et al. (2015) Genetics 199, 1-15.
- Cong, L. et al. (2013) Science 339, 819-23.
- Mali, P. et al. (2013) Science 339, 823-6.
- Li, D. et al. (2013) Nat Biotechnol 31, 681-3.
- Shen, B. et al. (2013) Cell Res 23, 720-3.
- Niu, Y. et al. (2014) Cell 156, 836-43.
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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For Research Use Only. Not For Use In Diagnostic Procedures.
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DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalStain is a trademark of Cell Signaling Technology, Inc.
Tween is a registered trademark of ICI Americas, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
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