Diagenode

CRISPR/Cas9 monoclonal antibody 4G10

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15200216-50
50 µg
$475.00
  Bulk order
Other format

Alternative name: Csn1

Monoclonal antibody raised in mouse against the N-terminus of the Cas9 nuclease (CRISPR-associated protein 9) using a recombinant protein.

Lot002
Concentration2.9 μg/μl
Species reactivityStreptococcus pyogenes
TypeMonoclonal
PurityProtein G purified
HostMouse
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution * References
Western Blotting 1:5,000 Fig 1
Immunoprecipitation 5 μg/IP Fig 2
Immunofluorescence 1:400 Fig 3
  • Validation data

    Western blot figure 1

    Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9
    Western blot was performed on protein extracts from HeLa cells transfected with Cas9 (lane 2) or from untransfected cells (lane 1) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200216), diluted 1:5,000 in PBS-T containing 0.5% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.

    Immunoprecipitation figure 2

    Figure 2. IP using the Diagenode monoclonal antibody directed against Cas9
    IP was performed on whole cell extracts (300 μg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 5 μg of the Diagenode antibody against Cas9 (Cat. No. C15200216). The immunoprecipitated proteins were subsequently analysed by Western blot with the polyclonal Cas9 antibody (Cat. No. C15310258, diluted 1:5,000). Lane 3 and 4 show the result of the IP, the input (15 μg) is shown in lane 1 and 2.

    Immunofluorescence figure 1

    Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9
    Hela cells were transiently transfected with a Cas9 expression vector. The cells were fixed with methanol/acetone and blocked in PBS containing 2% BSA. The cells were stained with the Cas9 antibody diluted 1;400, followed by incubation with an anti mouse secondary antibody coupled to AF488 for 1 h at RT (left). Nuclei were counter-stained with Hoechst 33342 (right).

  • Background

    CRISPR systems are adaptable immune mechanisms which are present in many bacteria to protect themselves from foreign nucleic acids, such as viruses, transposable elements or plasmids. Recently, the CRISPR/Cas9 (CRISPR-associated protein 9 nuclease, UniProtKB/Swiss-Prot entry Q99ZW2) system from S. pyogenes has been adapted for inducing sequence-specific double stranded breaks and targeted genome editing. This system is unique and flexible due to its dependence on RNA as the moiety that targets the nuclease to a desired DNA sequence and can be used to induce indel mutations, specific sequence replacements or insertions and large deletions or genomic rearrangements at any desired location in the genome. In addition, Cas9 can also be used to mediate upregulation of specific endogenous genes or to alter histone modifications or DNA methylation.

  • Testimonials

    When doing CRISPR experiments, it is essential to verify the expression levels of the Cas9 to ensure high-quality results. Diagenode's new CRISPR/Cas9 4G10 antibody is extremely specific, able to recognize different Cas9 fusion proteins (even those used for CRISPRi experiments) and compatible with many biochemical assays, such as immunofluorescence, western blot and immunoprecipitation. Among the different antibodies in the market, Diagenode's 4G10 CRISPR/Cas9 is our antibody of choice.

    Researcher at EPFL, Lausanne, Switzerland
  • Applications
    WB
    Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more
    IF
    Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more
    IP
    Immunoprecipitation Read more
  • Documents
    Datasheet CRISPR Cas9 4G10 monoclonal antibody DATASHEET
    Monoclonal antibody raised in mouse against the Cas9 nuclease (CRISPR-associated protein 9).
    Download
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
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    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
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  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: CRISPR/Cas9 monoclonal antibody 4G10 (Diagenode Cat# C15200216-50 Lot# 002). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Enhanced CRISPR/Cas9-mediated precise genome editing by improved design and delivery of gRNA, Cas9 nuclease, and donor DNA
    Liang W. et al.
    While CRISPR-based gene knock out in mammalian cells has proven to be very efficient, precise insertion of genetic elements via the cellular homology directed repair (HDR) pathway remains a rate-limiting step to seamless genome editing. Under the conditions described here, we achieved up to 56% targeted integration ...

    CRISPR-Mediated Gene Targeting of Human Induced Pluripotent Stem Cells
    Susan M. Byrne, George M. Church
    CRISPR/Cas9 nuclease systems can create double-stranded DNA breaks at specific sequences to efficiently and precisely disrupt, excise, mutate, insert, or replace genes. However, human embryonic stem cells and induced pluripotent stem cells (iPSCs) are more difficult to transfect and less resilient to DNA damage than...

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