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
CRISPR/Cas9 Antibody 4G10 (sample size)
Alternative name: Csn1
Monoclonal antibody raised in mouse against the Cas9 nuclease (CRISPR-associated protein 9).
| Lot | 003 |
|---|---|
| Concentration | 2 μg/μl |
| Species reactivity | Streptococcus pyogenes |
| Type | Monoclonal |
| Purity | Protein G purified monoclonal antibody in PBS containing 0.05 % Na-azide. |
| Host | Mouse |
| Storage Conditions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Precautions | This 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
Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9
Western blot was performed on protein extracts from HEK293T cells transfected with Cas9 (lane 1) or from untransfected cells (lane 2) 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.
Figure 2. IP using the Diagenode monoclonal antibody directed against CRISPR/Cas9
IP was performed on whole cell extracts from HEK293T cells transfected with a Cas9 expression vector (lane 1, 3 and 5), or untransfected cells (lane 2 and 4) using 5 µg of the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200216, lane 3 and 4) or with an equal amount of IgG, used as a negative control (lane 5). The immunoprecipitated proteins were subsequently analysed by Western blot with the polyclonal Cas9 antibody (Cat. No. C15310258, diluted 1:5,000). Lane 1 and 2 show the result of the input.
Figure 3. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9
HEK293T cells were transiently transfected with a Cas9 expression vector. The cells were fixed with 4% formaldehyde, permeabilized in 0,1% Triton X-100 and blocked in PBS containing 5% BSA. The cells were stained with the Cas9 antibody diluted 1;400 at 4°C o/n, followed by incubation with an anti mouse secondary antibody coupled to AF596 for 1 h at RT (left). Nuclei were counter-stained with DAPI (middle). A merge of the two stainings is shown on the right. - 出版物
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