Diagenode

CRISPR/Cas9 monoclonal antibody 7A9

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15200203
50 μg
$475.00
  Bulk order

Protein A purified monoclonal antibody in PBS containing 0.02 % Na-azide.

Lot003
Concentration1.9 µg/µl
Species reactivityStreptococcus pyogenes
TypeMonoclonal
PurityProtein A purified
HostMouse
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
Western Blotting 1:1,000 - 1:5,000 Fig 1, 2
Immunoprecipitation 1:200 Fig 3
Immunofluorescence 1:200 Fig 4
  • Validation Data

    WB

    Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against Cas9
    Western blot was performed on protein extracts from HEK293 cells transfected with a myc-tagged Cas9 using the Diagenode antibody against Cas9 (cat. No. C15200203). The antibody was used at different dilutions. The marker is shown on the left, position of the myc-tagged Cas9 protein is indicated on the right.

    WB

    Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against Cas9
    Western blot was performed on protein extracts from HEK293 cells transfected with a myc-tagged Cas9. Figure 2 shows the result of the WB on protein extracts from transfected (lane 2 and 4) and untransfected (lane 1 and 3) cells using the Diagenode antibody against Cas9 (Cat. No. C15200203, lane 1 and 2) or an anti-myc antibody (lane 3 and 4). The marker lane (M) was incubated with the Diagenode Blue ladder - HRP antibody (Cat. No. C15200202).

    WB

    Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9
    IP was performed on whole cell extracts (100 μg) from HEK293 cells transfected with a Flag-tagged Cas9 using the Diagenode antibody against Cas9 (cat. No. C15200203). The immunoprecipitated proteins were subsequently analysed by Western blot with the antibody. Lane 3 and 4 show the result of the IP; a negative IP control (IP on untransfected cells) and the input (15 μg) are shown in lane 2 and 1, respectively.

    WB

    Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against Cas9
    Hela cells were transiently transfected with a Flag-tagged Cas9 expression vector. 48 hours post transfection the cells were fixed in 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA for 2 hours at RT. The cells were stained with the Cas9 (fig 4A and B) or with an anti- Flag (fig 4C) antibody at 4°C o/n, followed by incubation with an anti mouse secondary antibody coupled to AF488 for 1 h at RT. Nuclei were counter-stained with Hoechst 33342 (bottom).

  • Background

    A breakthrough in genome engineering! The CRISPR/Cas9 (CRISPR-associated protein 9 nuclease) system uses a RNA-guided endonuclease technology which allows for inducing 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 up- or downregulation of specific endogenous genes or to alter histone modifications or DNA methylation.

  • Testimonials

    The antibody worked very well, even during our first attempt. We did a 1 hour incubation on a shaker at 4 degrees instead of overnight. We wanted to verify expression of Cas9 in various cell lines I had made (immunofluorescence). We have used the antibody a number of times and it works every time.

    Researcher at Harvard University
  • 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
    IP
    Immunoprecipitation 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
  • Documents
    Datasheet CRISPR/Cas9 monoclonal antibody DATASHEET
    This antibody has been raised against the N-terminal region of the CRISP/Cas9 protein.
    Download
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
    Download
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
    Download
  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: CRISPR/Cas9 monoclonal antibody 7A9 (Diagenode Cat# C15200203 Lot# 003). 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...

    A geminivirus-based guide RNA delivery system for CRISPR/Cas9 mediated plant genome editing
    Yin K, Han T, Liu G, Chen T, Wang Y, Yu AY, Liu Y
    CRISPR/Cas has emerged as potent genome editing technology and has successfully been applied in many organisms, including several plant species. However, delivery of genome editing reagents remains a challenge in plants. Here, we report a virus-based guide RNA (gRNA) delivery system for CRISPR/Cas9 mediated plant ge...

    Rapid and highly efficient mammalian cell engineering via Cas9 protein transfection.
    Liang X, Potter J, Kumar S, Zou Y, Quintanilla R, Sridharan M, Carte J, Chen W, Roark N, Ranganathan S, Ravinder N, Chesnut JD
    CRISPR-Cas9 systems provide a platform for high efficiency genome editing that are enabling innovative applications of mammalian cell engineering. However, the delivery of Cas9 and synthesis of guide RNA (gRNA) remain as steps that can limit overall efficiency and ease of use. Here we describe methods for rapid synt...

    A localized nucleolar DNA damage response facilitates recruitment of the homology-directed repair machinery independent of cell cycle stage.
    van Sluis M, McStay B
    DNA double-strand breaks (DSBs) are repaired by two main pathways: nonhomologous end-joining and homologous recombination (HR). Repair pathway choice is thought to be determined by cell cycle timing and chromatin context. Nucleoli, prominent nuclear subdomains and sites of ribosome biogenesis, form around nucleolar ...

    Genome-wide CRISPR screen in a mouse model of tumor growth and metastasis.
    Chen S, Sanjana NE, Zheng K, Shalem O, Lee K, Shi X, Scott DA, Song J, Pan JQ, Weissleder R, Lee H, Zhang F, Sharp PA
    Genetic screens are powerful tools for identifying genes responsible for diverse phenotypes. Here we describe a genome-wide CRISPR/Cas9-mediated loss-of-function screen in tumor growth and metastasis. We mutagenized a non-metastatic mouse cancer cell line using a genome-scale library with 67,405 single-guide RNAs (s...

    Use of the CRISPR/Cas9 system as an intracellular defense against HIV-1 infection in human cells.
    Liao HK, Gu Y, Diaz A, Marlett J, Takahashi Y, Li M, Suzuki K, Xu R, Hishida T, Chang CJ, Esteban CR, Young J, Izpisua Belmonte JC
    To combat hostile viruses, bacteria and archaea have evolved a unique antiviral defense system composed of clustered regularly interspaced short palindromic repeats (CRISPRs), together with CRISPR-associated genes (Cas). The CRISPR/Cas9 system develops an adaptive immune resistance to foreign plasmids and viruses by...

    An inducible lentiviral guide RNA platform enables the identification of tumor-essential genes and tumor-promoting mutations in vivo.
    Aubrey BJ, Kelly GL, Kueh AJ, Brennan MS, O'Connor L, Milla L, Wilcox S, Tai L, Strasser A, Herold MJ
    The CRISPR/Cas9 technology enables the introduction of genomic alterations into almost any organism; however, systems for efficient and inducible gene modification have been lacking, especially for deletion of essential genes. Here, we describe a drug-inducible small guide RNA (sgRNA) vector system allowing for ubiq...

    An inducible lentiviral guide RNA platform enables the identification of tumor-essential genes and tumor-promoting mutations in vivo
    Aubrey BJ et al.
    The CRISPR/Cas9 technology enables the introduction of genomic alterations into almost any organism; however, systems for efficient and inducible gene modification have been lacking, especially for deletion of essential genes. Here, we describe a drug-inducible small guide RNA (sgRNA) vector system allowing for ubiq...

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