CRISPR/Cas9 monoclonal antibody 4G10

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

CRISPR/Cas9 Antibody for Western blot

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.

CRISPR/Cas9 Antibody validated in IP

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.

CRISPR/Cas9 Antibody for IF

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.

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

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

Datasheet CRISPR Cas9 4G10 monoclonal antibody DATASHEET Monoclonal antibody raised in mouse against the Cas9 nuclease (CRISPR-associated protein 9).	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
Accurate QC to optimize CRISPR/Cas9 genome editing specificity POSTER The CRISPR/Cas9 technology is delivering superior genetic models for fundamental disease res...	Download

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Cis-regulatory interfaces reveal the molecular mechanisms underlying the notochord gene regulatory network of Ciona
Negrón-Piñeiro L. J. et al.
Tissue-specific gene expression is fundamental in development and evolution, and is mediated by transcription factors and by the cis-regulatory regions (enhancers) that they control. Transcription factors and their respective tissue-specific enhancers are essential components of gene regulatory networks respons...

Monitoring autochthonous lung tumors induced by somatic CRISPR geneediting in mice using a secreted luciferase.
Merle N. et al.
BACKGROUND: In vivo gene editing of somatic cells with CRISPR nucleases has facilitated the generation of autochthonous mouse tumors, which are initiated by genetic alterations relevant to the human disease and progress along a natural timeline as in patients. However, the long and variable, orthotopic tumor growth ...

Systematic comparison of CRISPR-based transcriptional activatorsuncovers gene-regulatory features of enhancer-promoter interactions.
Wang K. et al.
Nuclease-inactivated CRISPR/Cas-based (dCas-based) systems have emerged as powerful technologies to synthetically reshape the human epigenome and gene expression. Despite the increasing adoption of these platforms, their relative potencies and mechanistic differences are incompletely characterized, particularly at h...

DNA Methylation Editing by CRISPR-guided Excision of 5-Methylcytosine.
Devesa-Guerra I, Morales-Ruiz T, Pérez-Roldán J, Parrilla-Doblas JT, Dorado-León M, García-Ortiz MV, Ariza RR, Roldán-Arjona T
Tools for actively targeted DNA demethylation are required to increase our knowledge about regulation and specific functions of this important epigenetic modification. DNA demethylation in mammals involves TET-mediated oxidation of 5-methylcytosine (5-meC), which may promote its replication-dependent dilution and/or...

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 ...

Optimization of CRISPR/Cas9 Delivery to Human Hematopoietic Stem and Progenitor Cells for Therapeutic Genomic Rearrangements.
Lattanzi A, Meneghini V, Pavani G, Amor F, Ramadier S, Felix T, Antoniani C, Masson C, Alibeu O, Lee C, Porteus MH, Bao G, Amendola M, Mavilio F, Miccio A
Editing the β-globin locus in hematopoietic stem cells is an alternative therapeutic approach for gene therapy of β-thalassemia and sickle cell disease. Using the CRISPR/Cas9 system, we genetically modified human hematopoietic stem and progenitor cells (HSPCs) to mimic the large rearrangements in the &beta...

Optimization of CRISPR/Cas9 genome editing for loss-of-function in the early chick embryo
Gandhi S. et al.
The advent of CRISPR/Cas9 has made genome editing possible in virtually any organism, including those not previously amenable to genetic manipulations. Here, we present an optimization of CRISPR/Cas9 for application to early avian embryos with improved efficiency via a three-fold strategy. First, we employed Cas9 pr...

Enhanced CRISPR/Cas9-mediated precise genome editing by improved design and delivery of gRNA, Cas9 nuclease, and donor DNA
Liang W. et al.
While CRISPR-based gene knock out in mammalian cells has proven to be very efficient, precise insertion of genetic elements via the cellular homology directed repair (HDR) pathway remains a rate-limiting step to seamless genome editing. Under the conditions described here, we achieved up to 56% targeted integration ...

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...

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CRISPR/Cas9 monoclonal antibody 4G10

How to properly cite our product/service in your work

iDeal ChIP-seq kit for Histones

MicroPlex Library Preparation Kit v2 (12 indices)

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