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10612
Cas9 (7A9-3A3) Mouse mAb (BSA and Azide Free)
Primary Antibodies
Monoclonal Antibody

Cas9 (7A9-3A3) Mouse mAb (BSA and Azide Free) #10612

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  1. WB
  2. IHC
  3. IF
  4. F
Western Blotting Image 1: Cas9 (7A9-3A3) Mouse mAb (BSA and Azide Free)
Western blot analysis of extracts from 293T cells, mock transfected (-), or transfected with a construct expressing Cas9 protein isolated from S.pyogenes (spCas9) (+), or recombinant nuclease-deficient Cas9 (dCas9) protien, or recombinant Cas9 nickase (niCas9) protein, using Cas9 (7A9-3A3) Mouse mAb. Data were generated using the standard formulation of this product.
Western Blotting Image 2: Cas9 (7A9-3A3) Mouse mAb (BSA and Azide Free)
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). Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: Cas9 (7A9-3A3) Mouse mAb (BSA and Azide Free)
Immunohistochemical analysis of Cas9 expression in Nprl2-deficient 293 (left, positive) and untreated 293 (right, negative) cell pellets using Cas9 (7A9-3A3) Mouse mAb. 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. Data were generated using the standard formulation of this product.
Immunofluorescence Image 1: Cas9 (7A9-3A3) Mouse mAb (BSA and Azide Free)
Confocal immunofluorescent analysis of Cas9 expression in Nprl2-deficient (left) and untreated 293 cells (right) using Cas9 (7A9-3A3) Mouse mAb (green). 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. Data were generated using the standard formulation of this product.
Flow Cytometry Image 1: Cas9 (7A9-3A3) Mouse mAb (BSA and Azide Free)
Flow cytometric analysis of HEK-293 cells untreated (blue) or transfected with NPRL2 knock out (-/-) plasmid (green) Cas9 (7A9-3A3) Mouse mAb. Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody. Data were generated using the standard formulation of this product.
To Purchase # 10612
Cat. # Size Qty. Price
10612SF
100 µg
$ 746

Supporting Data

REACTIVITY All
SENSITIVITY Transfected Only
MW (kDa) 160
Source/Isotype Mouse IgG1

Application Key:

  • WB-Western Blot
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • C&R-CUT&RUN
  • C&T-CUT&Tag
  • DB-Dot Blot
  • eCLIP-eCLIP
  • IF-Immunofluorescence
  • F-Flow Cytometry

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Vir-Virus
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Usage Information

This product is the carrier free version of product #14697. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN, or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

Formulation

Supplied in 1X PBS, BSA and Azide Free.

For standard formulation of this product see product #14697.

Storage

Store at –20°C. This product will freeze at –20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity / Sensitivity

Cas9 (7A9-2A3) Mouse mAb (BSA and Azide Free) recognizes transfected levels of total Cas9 protein.

Species Reactivity:

All Species Expected

Source / Purification

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

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.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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