Diagenode

Antibodies

CRISPR/Cas9 antibodies

Best CRISPR/Cas9 antibodies on the market!

Diagenode offers a broad range of highly validated CRISPR/Cas9 antibodies, recommended for different applications (WB, IF, IP and ChIP). These antibodies have been raised against the N- or C-terminus of the Cas9 nuclease from Streptococcus pyogenes or Staphylococcus aureus.

  • S. aureus CRISPR/Cas9 antibodies NEW!
  • S. pyogenes CRISPR/Cas9 antibodies

    The CRISPR/Cas9 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.

    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.

    Learn more

    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).

    Learn more

    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.

    Learn more

    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

Four critical reasons to use anti-CRISPR/Cas9 antibodies

CRISPR/Cas9 genome editing allows for double-stranded DNA breaks at specific sequences to efficiently disrupt, excise, mutate, insert, or replace genes. The precision of transfection and the level of Cas9 expression should be controlled during the editing processes using specific anti-CRISPR/Cas9 antibodies.

Reason #1 Check the transfection success of the Cas9 protein

Check Cas9 expression level using Western blot with validated anti-Cas9 antibodies.

Reason #2 Check that the Cas9 protein was delivered to the nucleus

Determine the nuclear localization of Cas9 protein via IF or IHC using validated anti-Cas9 antibodies

Reason #3 Check the Cas9 expression level

Transient systems: Check that Cas9 expression was transient by using Western blot and anti-Cas9 antibodies. Prolonged Cas9 expression will lead to more off-target mutations.

Stable transformants: Isolate several clones and screen to check the level of Cas9 expression using Western blot and specific anti-Cas9 antibodies. High level of Cas9 expression can lead to non-specific activity.

Reason #4 Check the binding specificity of Cas9

Test the binding specificity with an sgRNA of choice by using the ChIP-grade anti-Cas9 antibody and the primers for targeted and non-targeted region to see if Cas9 is bound to the correct region.

Products

ProductApplicationPurity
CRISPR/Cas9 C-terminal monoclonal antibody
(50 µg)
IF IP WB Protein G purified
CRISPR/Cas9 monoclonal antibody
(50 μg/25 μl)
ChIP-qPCR IF IP WB Protein G purified
CRISPR/Cas9 monoclonal antibody 4G10
(50 µg)
IF IP WB Protein G purified
CRISPR/Cas9 polyclonal antibody
(100 μl)
ChIP-qPCR IF IP WB Whole antiserum
CRISPR/Cas9 - HRP monoclonal antibody 7A9
(500 µl)
WB Protein G purified
CRISPR/Cas9 C-terminal monoclonal antibody
(50 μg/23 μl)
ChIP-qPCR IF WB Protein G purified
CRISPR/Cas9 monoclonal antibody 7A9
(50 μg)
IF IP WB Protein A purified

S. aureus CRISPR/Cas9 antibodies

ProductApplicationPurity
S. aureus CRISPR/Cas9 monoclonal antibody
(50 μg/ 25 μl)
IF IP WB Protein G purified
S. aureus CRISPR/Cas9 polyclonal antibody (C-terminal)
(100 μl)
IF IP WB Whole antiserum
S. aureus CRISPR/Cas9 polyclonal antibody (N-terminal)
(100 μl)
IF IP WB Whole antiserum

Events

  • Transcription and Chromatin
    EMBL Heidelberg
    Aug 27-Aug 30, 2016
 See all events

Twitter feed

News

 See all news