Diagenode

CRISPR/Cas9 monoclonal antibody (sample size)

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15200229-10
10 µg
$99.00
  Bulk order
Other format

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

Lot001
Concentration2 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
ChIP * 5 μg/ChIP Fig 1
Western Blotting 1:1,000 - 1:10,000 Fig 2
Immunoprecipitation 6 μg/IP Fig 3
Immunofluorescence 1:400 Fig 4
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 μg per IP.
  • Validation Data

    ChIP figure 1

    Figure 1. ChIP using the Diagenode monoclonal antibody directed against Cas9
    ChIP was performed on NIH3T3 cells stably expressing GFP-H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50 μg chromatin was incubated overnight at 4°C with 5 or 10 μg of either an anti-FLAG antibody or the Diagenode antibody against Cas9 (Cat. No. C15200229). Mouse IgG was used as a negative IP control. qPCR was performed with primers specific for the GFP gene, and for a non-targeted region (protein kinase C delta, Prkcd), used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    WB figure 2 AWB figure 2 B

    Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9
    Figure 1A: Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200229). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. Figure 1B: Western blot was performed on protein extracts from HeLa cells transfected with Cas9 (lane 1) or from untransfected cells (lane 2) using the Diagenode antibody against CRISPR/Cas9 (Cat. No. C15200229), diluted 1:4,000 in PBS-T containing 3% NFDM. The marker is shown on the left, position of the Cas9 protein is indicated on the right.

    IP figure 3

    Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9
    IP was performed on whole cell extracts (300 μg) from HEK293 cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 6 μg of the Diagenode antibody against Cas9 (Cat. No. C15200229). The immunoprecipitated proteins were subsequently analysed by Western blot with the polyclonal Cas9 antibody (Cat. No. C15310258, diluted 1:8,000). Lane 3 and 4 show the result of the IP, the input (15 μg) is shown in lane 1 and 2.

    IF figure 4

    Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9
    HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.

  • 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
    ChIP-qPCR (ab)
    Read more
  • Documents
    CRISPR/Cas9 monoclonal antibody C15200229 DATASHEET
    CRISPR/Cas9 monoclonal antibody datasheet
    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 (sample size) (Diagenode Cat# C15200229-10 Lot# 001). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    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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