Diagenode

CRISPR/Cas9 antibodies

S. pyogenes CRISPR/Cas9 antibodies

Diagenode offers the broad range of antibodies raised against the N- or C-terminus of the Cas9 nuclease from Streptococcus pyogenes. These highly specific polyclonal and monoclonal antibodies are validated in Western blot, immunoprecipitation, immunofluorescence and in chromatin immunoprecipitation.

S. pyogenes CRISPR/Cas9 antibodies

Discover our first monoclonal CRISPR/Cas9 antibody validated in ChIP

  • Validated in chromatin immunoprecipitation
  • Performs better than FLAG antibody
  • Excellent for WB, IF and IP

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).

First ChIP-grade CRISPR/Cas9 polyclonal antibody

  • Excellent polyclonal antibody for chromatin immunoprecipitation
  • Optimized for highest ChIP specificity and yields
  • Validated for all applications including immunoblotting, immunofluorescence and western blot

ChIP was performed on NIH3T3 cells stably expressing GFP- H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50 μg chromatin was incubated with either 5 μg of an anti-FLAG antibody or 2 μl of the Diagenode antibody against Cas9. The pre-immune serum (PPI) was used as negative IP control. Then qPCR was performed with primers specific for the GFP gene, and for two non-targeted regions: Ppap2c and Prkcd, used as negative controls. This figure shows the recovery, expressed as a % of input.

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CRISPR/Cas9 monoclonal antibody 4G10

  • Antibody raised against N-terminus of Cas9 nuclease
  • Validated for western blot, IP and immunofluorescence

Immunofluorescence: Hela cells were transiently transfected with a Cas9 expression vector. The cells were fixed in 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA. The cells were stained with the Cas9 antibody at 4°C o/n, followed by incubation with an anti mouse secondary antibody coupled to AF488 for 1 h at RT. Nuclei were counter-stained with Hoechst 33342 (right).

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CRISPR/Cas9 C-terminal monoclonal antibody

  • Antibody raised against C-terminus of Cas9 nuclease
  • Validated for western blot, IP and immunofluorescence

Western blot was performed on 20 μg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9. The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right.

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Which CRISPR/Cas9 antibody is the best for your application?

Antibody WB IF IP ChIP Antibody raised against
CRISPR/Cas9 monoclonal antibody +++ +++ ++ +++ N-terminus of Cas9 nuclease
CRISPR/Cas9 polyclonal antibody ++ ++ ++ +++ N-terminus of Cas9 nuclease
CRISPR/Cas9 monoclonal antibody 4G10 +++ +++ ++ no N-terminus of Cas9 nuclease
CRISPR/Cas9 C-terminal monoclonal antibody NEW! +++ +++ no + C-terminus of Cas9 nuclease
CRISPR/Cas9 C-terminal monoclonal antibody ++ ++ + no C-terminus of Cas9 nuclease
CRISPR/Cas9 monoclonal antibody 7A9 ++ ++ ++ no N-terminus of Cas9 nuclease
CRISPR/Cas9 - HRP monoclonal antibody 7A9 +++ no no no N-terminus of Cas9 nuclease

Products

Cat. No.ProductApplicationPurity
C15200223-50 CRISPR/Cas9 C-terminal monoclonal antibody
(50 µg)
IF IP WB Protein G purified
C15200229 CRISPR/Cas9 monoclonal antibody
(50 μg/25 μl)
ChIP-qPCR IF IP WB Protein G purified
C15200216-50 CRISPR/Cas9 monoclonal antibody 4G10
(50 µg /25 µl)
IF IP WB Protein G purified monoclonal antibody in PBS containing 0.05 % Na-azide.
C15310258 CRISPR/Cas9 polyclonal antibody
(100 μl)
ChIP-qPCR IF IP WB Whole antiserum
C15200215 CRISPR/Cas9 - HRP monoclonal antibody 7A9
(500 µl)
WB Protein G purified
C15200231-50 CRISPR/Cas9 C-terminal monoclonal antibody
(50 μg/23 μl)
ChIP-qPCR IF WB Protein G purified
C15200203 CRISPR/Cas9 monoclonal antibody
(50 µg/17 µl)
IF IP WB Protein A purified monoclonal antibody in PBS containing 0.02 % Na-azide.

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