#19526 Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb
IHC-P: 抗体希釈液 / 抗原賦活化
|Cas9 (S. pyogenes) (E7M1H) Rabbit mAb recognizes transfected levels of total Cas9 (S. pyogenes) protein. This antibody does not cross-react with Cas9 (S. aureus), FnCpf1, and AsCpf1 proteins.|
|Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu833 of Cas9 (S. Pyogenes) protein.|
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Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Cas9 (S. pyogenes), using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (upper) and Myc-Tag (71D10) Rabbit mAb #2278 (lower).
Immunohistochemical analysis of Cas9 (S. pyogenes) expression in paraffin-embedded Nprl2-deficient 293 cell pellet (left, positive) or untreated 293 cell pellet (right, negative) using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb. Nprl2 expression was knocked out in the Nprl2-deficient cells by transient transfection of Cas9 (S. pyogenes) and Nprl2-specific guide sequences. 293/Nrpl2 -/- cells were kindly provided by Rachel Wolfson, Lynne Chantranupong, and David Sabatini of MIT, Cambridge, MA.
Immunohistochemical analysis of Cas9 expression in paraffin-embedded Cas9 (S. Pyogenes) transgenic mouse brain (top-left, positive) and wild type mouse tissues (negative): brain (top-center), colon (top-right), liver (bottom-left), lung (bottom-center), and pancreas (bottom-right), using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb.
Confocal immunofluorescent analysis of brain from constitutive Cas9 (S. pyogenes)-expressing mice (left, positive) or wild type mice (right, negative), using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Confocal immunofluorescent analysis of 293T cells transiently transfected with a myc-tagged Cas9 (S. pyogenes) construct, using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (green) and Myc-Tag (9B11) Mouse mAb #2276 (red). Colocalization of green and red signals appear yellow in the composite image (bottom-right). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow cytometric analysis of 293 cells, untransfected (blue) or transfected with Cas9 (S. pyogenes) (green), using Cas9 (S. pyogenes) (E7M1H) XP® Rabbit mAb (solid lines) or concentraion-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-Rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as secondary antibody.
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).
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