CRISPR/Cas9 Antibody
Monoclonal antibody raised in mouse against the N-terminus of the Cas9 nuclease (CRISPR-associated protein 9) using a recombinant protein.
| Lot | 002 |
|---|---|
| Concentration | 2 μg/μl |
| Species reactivity | Streptococcus pyogenes |
| Type | Monoclonal |
| Purity | Protein G purified |
| Host | Mouse |
| Precautions | This 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
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).
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.
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.
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 N-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.
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 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.
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
| Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar... | Download |
| Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e... | Download |
| CRISPR/Cas9 monoclonal antibody C15200229 DATASHEET CRISPR/Cas9 monoclonal antibody datasheet | Download |
| Accurate QC to optimize CRISPR/Cas9 genome editing specificity POSTER The CRISPR/Cas9 technology is delivering superior genetic models for fundamental disease res... | Download |
Safety sheets
| CRISPR/Cas9 Antibody SDS US en | Download |
| CRISPR/Cas9 Antibody SDS GB en | Download |
| CRISPR/Cas9 Antibody SDS FR fr | Download |
| CRISPR/Cas9 Antibody SDS BE fr | Download |
| CRISPR/Cas9 Antibody SDS ES es | Download |
| CRISPR/Cas9 Antibody SDS JP ja | Download |
| CRISPR/Cas9 Antibody SDS DE de | Download |
| CRISPR/Cas9 Antibody SDS BE nl | Download |
Publications
How to properly cite our product/service in your workWe strongly recommend using this: CRISPR/Cas9 Antibody (Hologic Diagenode Cat# C15200229-100 Lot# 002). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
TurboCas: A method for locus-specific labeling of genomic regions and isolating their associated protein interactome |
Genome-wide CRISPR screen identifies protein pathways modulating tauprotein levels in neurons |
Transgenic mice for in vivo epigenome editing with CRISPR-based systems |
Circuit-specific hippocampal ΔFosB underlies resilience to
stress-induced social avoidance. |
Endogenous retroviruses are a source of enhancers with oncogenic potential in acute myeloid leukaemia |
Improved double-nicking strategies for COL7A1 editing by homologous recombination |
Improved Double-Nicking Strategies for COL7A1-Editing by Homologous Recombination. |
Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies. |
RNA-Based dCas9–VP64 System Improves the Viability of Cryopreserved Mammalian Cells |
CRISPR-Mediated Gene Targeting of Human Induced Pluripotent Stem Cells |