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.
Diagenode offers the best antibody against CRISPR/Cas9 on the market! It has been raised against Cas9 from Streptococcus pyogenes and gives excellent results in all major applications (immunoblot, immunoprecipitation and immunofluorescence). The perfect tool for monitoring your CRISPR/Cas9 experiments.
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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
HeLa cells were transfected with Cas9 and stained with the CRISPR/Cas9 antibody 4G10 at 4°C overnight, followed by incubation with an anti-mouse secondary antibody coupled to AF488 for 1 hour at room temperature (upper figure). The lower figure shows counter-staining of the nuclei with Hoechst 33342.
Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using antibodies against Cas9. The antibodies were used at different dilutions (from left to right 1:250, 1:500, 1:1,000 and 1:2,000). The results obtained with the 7A9 clone are shown on the left, the results obtained with the new 4G10 clone are shown on the right.
Extremely specific and able to recognize different Cas9 fusion proteinsWestern blot was performed on protein extracts from HeLa cells transfected with dCas9 or dCas9-Krab, using Diagenode’s CRISPR/Cas9 4G10 antibody. 1 million cells were transfected with 1 µg of Cas9 plasmids. The obtained signal is in the expected range. Lane 1 and 2: transfected cells with low levels of dCas9. Lane 3: wild-type as negative control. Lane 4: positive control: selected cell line expressing high levels of Cas9 wild type (not the dead-Cas9). (Data kindly provided by Patricia Renck Nunes, EPFL, Lausanne, Switzerland)