CRISPR/Cas9 Antibody (sample size)

Catalog Number
20 μl
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Polyclonal antibody raised in rabbit against the Cas9 nuclease (CRISPR-associated protein 9) using a recombinant protein. 

ConcentrationNot determined
Species reactivityStreptococcus pyogenes
TypePolyclonal, ChIP grade, ChIP-seq grade
PurityWhole antiserum from rabbit containing 0.05% azide.
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP/ChIP-seq * 2-5 µl/ChIP Fig 1, 2
Western Blotting 1:5,000 Fig 3
Immunoprecipitation 1 µl/IP Fig 4
Immunofluorescence 1:1,000 Fig 5

* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µl per IP.


I used ChIP-qPCR with the Diagenode CRISPR/Cas9 polyclonal antibody to successfully show that Cas9 binds to the target region of my sgRNA, validating my CRISPR experiment. The antibody produced minimal background signal at non-specific genomic regions. I am now using the antibody to validate further sgRNA in different CRISPR cell lines.

ChIP was performed on Jurkat cells expressing dCas9-VP64-mCherry and a sgRNA targeting the IL1RN promoter. Each IP was performed using 4 million cells and 2 µL CRISPR/Cas9 polyclonal antibody (Diagenode C15310258) or 1 µg rabbit IgG control antibody (Diagenode C15410206). qPCR was carried out on undiluted ChIP DNA using SYBR green and PCR primers directed against the sgRNA binding site at IL1RN, as well as two non-target regions at the SLC4A1 and TP53 promoters. ChIP enrichment was measured using the percent input method.

Researcher from the University of Manchester
  • Validation data

    CRISPR/Cas9 Antibody ChIP Grade

    Figure 1. ChIP using the Diagenode antibody directed against Cas9
    ChIP was performed on NIH3T3 cells stably expressing GFP-H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50μg chromatin was incubated overnight at 4°C with either 5 μg of an anti-FLAG antibody or 2 μl of the Diagenode antibody against Cas9 (cat. No. C15310258). The pre-immune serum (Cas9, PPI) was used as negative IP control. qPCR was performed with primers specific for the GFP gene, and for two non-targeted regions phosphatidic acid phosphatase type 2C (Ppap2c) and protein kinase C delta (Prkcd), used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    CRISPR/Cas9 Antibody Validation in WB

    Figure 2. Western blot analysis using the Diagenode antibody directed against Cas9
    Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310258). The antibody was diluted 1:1,000 (lane 2) or 1:5,000 (lane 3). Lane 1 shows the result with the pre-immune serum. The marker is shown on the left, the position of the Cas9 protein is indicated on the right.

    CRISPR/Cas9 Antibody Validation in IP

    Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9
    IP was performed on whole cell extracts (500 μg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 μl of the Diagenode antibody against Cas9 (cat. No. C15310258). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (25 μg) is shown in lane 1 and 2.

    CRISPR/Cas9 Antibody Validation in IF

    Figure 4. Immunofluorescence using the Diagenode antibody directed against Cas9
    HeLa cells expressing Cas9 under the control of the tight TRE promoter 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 antibody (cat. No. C15310258) diluted 1:1000, followed by incubation with a goat anti-rabbit secondary antibody coupled to AF594. Nuclei were counterstained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline.

  • Target Description

    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 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

    Diagenode’s CRISPR/Cas9 polyclonal antibody shows superior signal than the original clone 7A9: a researcher from EPFL in Lausanne, Switzerland has compared these two antibodies in Western blot.

    Western blot was performed using HCT116 DKO cells transduced with Krab-dCas9 (2) or non-transduced (1) cells. Then, 100,000 cells were lysed in sample buffer 2x and boiled 5 min at 95°C before loading in a 15% acrylamide gel. The same sample was loaded 3x in the same gel. The membrane was cut in 3 parts for each antibody. Membrane was blocked 1h with 3% BSA at RT. Antibodies were diluted 1:1,000 in 3% BSA and incubated overnight at 4°C. Secondary incubation was done for 1h at RT (1:10,000 dilution). Anti-hnRNPA1 was used as a loading control.

    EPFL in Lausanne
  •  Applications
    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
    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
    Immunoprecipitation Read more
  •  Documents
    C15310258-Datasheet CRISPR Cas9 Polyclonal antibody DATASHEET
    Datasheet description
    Accurate QC to optimize CRISPR/Cas9 genome editing specificity POSTER
    The CRISPR/Cas9 technology is delivering superior genetic models for fundamental disease res...
    Optimize the selection of guide RNA by ChIP to keep CRISPR on-target APPLICATION NOTE
    The mechanisms of target recognition and target specificity of the Cas9 protein is still not comp...
  •  Safety sheets
    CRISPR/Cas9 Antibody - ChIP-seq Grade SDS US en Download
    CRISPR/Cas9 Antibody - ChIP-seq Grade SDS GB en Download
    CRISPR/Cas9 Antibody - ChIP-seq Grade SDS BE fr Download
    CRISPR/Cas9 Antibody - ChIP-seq Grade SDS FR fr Download
    CRISPR/Cas9 Antibody - ChIP-seq Grade SDS ES es Download
    CRISPR/Cas9 Antibody - ChIP-seq Grade SDS DE de Download
    CRISPR/Cas9 Antibody - ChIP-seq Grade SDS JP ja Download
    CRISPR/Cas9 Antibody - ChIP-seq Grade SDS BE nl Download
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: CRISPR/Cas9 Antibody (sample size) (Diagenode Cat# C15310258-20 Lot# A2508-004). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    TGFβ promotes widespread enhancer chromatin opening and operates ongenomic regulatory domains.
    Guerrero-Martínez J. et al.
    The Transforming Growth Factor-β (TGFβ) signaling pathway controls transcription by regulating enhancer activity. How TGFβ-regulated enhancers are selected and what chromatin changes are associated with TGFβ-dependent enhancers regulation are still unclear. Here we report that TGFβ treatment...

    A gene therapy for inherited blindness using dCas9-VPR–mediatedtranscriptional activation
    Böhm, Sybille and Splith, Victoria and Riedmayr, Lisa Maria and Rötzer,René Dominik and Gasparoni, Gilles and Nordström, Karl J. V. and Wagner,Johanna Elisabeth and Hinrichsmeyer, Klara Sonnie and Walter, Jörn andWahl-Schott, Christian and Fenske, Stef
    Catalytically inactive dCas9 fused to transcriptional activators (dCas9-VPR) enables activation of silent genes. Many disease genes have counterparts, which serve similar functions but are expressed in distinct cell types. One attractive option to compensate for the missing function of a defective gene could be to t...

    Guidelines for optimized gene knockout using CRISPR/Cas9
    Campenhout CV et al.
    CRISPR/Cas9 technology has evolved as the most powerful approach to generate genetic models both for fundamental and preclinical research. Despite its apparent simplicity, the outcome of a genome-editing experiment can be substantially impacted by technical parameters and biological considerations. Here, we present ...

    CRISPR-mediated activation of a promoter or enhancer rescues obesity caused by haploinsufficiency.
    Matharu N, Rattanasopha S, Tamura S, Maliskova L, Wang Y, Bernard A, Hardin A, Eckalbar WL, Vaisse C, Ahituv N
    A wide range of human diseases result from haploinsufficiency, where the function of one of the two gene copies is lost. Here, we targeted the remaining functional copy of a haploinsufficient gene using CRISPR-mediated activation (CRISPRa) in and heterozygous mouse models to rescue their obesity phenotype. Transgeni...

    (Po)STAC (Polycistronic SunTAg modified CRISPR) enables live-cell and fixed-cell super-resolution imaging of multiple genes
    Neguembor M.V. et al.
    CRISPR/dCas9-based labeling has allowed direct visualization of genomic regions in living cells. However, poor labeling efficiency and signal-to-background ratio have limited its application to visualize genome organization using super-resolution microscopy. We developed (Po)STAC (Polycistronic SunTAg modified CRISP...


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