CRISPR-Cas9 Antibody (7A9-3A3) [DyLight 650]

Novus Biologicals | Catalog # NBP2-36440C

Novus Biologicals
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Key Product Details

Species Reactivity

Mouse, Bacteria, Bovine, Chicken, Fungi

Applications

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry Whole-Mount, Western Blot, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Label

DyLight 650 (Excitation = 652 nm, Emission = 672 nm)

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 7A9-3A3
View all CRISPR-Cas9 Primary Antibodies »

Product Specifications

Immunogen

This CRISPR-Cas9 antibody (7A9-3A3) - N-Terminus was raised against Recombinant Cas9 within the N-terminal region of Streptococcus pyogene. [Uniprot: Q99ZW2].

Specificity

This CRISPR-Cas9 antibody (7A9-3A3) - N-Terminus is specific toCas9 protein from Streptococcus pyogene serotype M1.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Applications for CRISPR-Cas9 Antibody (7A9-3A3) [DyLight 650]

Application
Recommended Usage

Immunoblotting

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry Whole-Mount

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry-Frozen

Optimal dilutions of this antibody should be experimentally determined.

Immunoprecipitation

Optimal dilutions of this antibody should be experimentally determined.

Knockout Validated

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
Optimal dilution of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

50mM Sodium Borate

Preservative

0.05% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C in the dark.

Background: CRISPR-Cas9

Clustered regularly interspaced short palindromic repeats (CRISPRs) are derived from DNA fragments of bacteriophages that infect prokaryotes. When infected, the bacteria capture snips of DNA from the invading virus to create CRISPR arrays. During subsequent infections, the bacteria produce RNA segments from the CRISPR arrays to target the virus' DNA. CRISPR-associated protein 9 (Cas9) is RNA-guided, binds DNA, and is a cleaving enzyme that functions as an integral component of the bacterial CRISPR adaptive immune system that targets the virus' DNA to disable it (1). To check for sites complementary to the 20 base pair spacer region of the guide RNA (gRNA) of the CRISPR, Cas9 unwinds foreign DNA that invades the bacteria. If the DNA substrate is complementary to the gRNA, Cas9 cleaves the invading DNA, rendering the virus disabled. The presence of a 5'-NGG-3' protospacer adjacent motif (PAM) sequence immediately downstream of the target DNA (protospacer) is required for Cas9 cleavage of foreign DNA. As PAM is absent in bacterial CRISPR loci, cleavage of the host genome is avoided and provides a novel sequence for identification of foreign DNA by Cas9.

Using CRISPR-Cas9 technology, double-stranded DNA breaks may be induced within specific targeted genome sequences (target DNA; protospacer) for insertion or removal of DNA sequences for gene editing applications. To target a specific loci, a gRNA that will bind to a specific target sequence of DNA within a genome is created. The gRNA will recognize the DNA sequence, and the Cas9 enzyme will cleave the DNA at the targeted location. Once the targeted DNA is removed by Cas9, the cell's own DNA repair mechanism is used to insert or remove a DNA sequence for genomic editing.

Cas9 detection is used to confirm and evaluate CRISPR Cas9 gRNA transfection efficiency. Western blot analysis of CRISPR-Cas9 gRNA transfected cell lysates with Cas9 antibodies identifies the protein having a theoretical molecular weight of 160kDa. Broad areas of research are benefiting from CRISPR-Cas9 based gene editing tools including studies of basic immunity functions, genetic screening and disease treatment (2). Ethical concerns have led to many countries making it illegal to manipulate human germline cells or perform embryo genome editing.

References

1. Oakes, B. L., Fellmann, C., Rishi, H., Taylor, K. L., Ren, S. M., Nadler, D. C.,... Savage, D. F. (2019). CRISPR-Cas9 Circular Permutants as Programmable Scaffolds for Genome Modification. Cell, 176(1-2), 254-267.e216. doi:10.1016/j.cell.2018.11.052

2. Chiou, S. H., Winters, I. P., Wang, J., Naranjo, S., Dudgeon, C., Tamburini, F. B.,... Winslow, M. M. (2015). Pancreatic cancer modeling using retrograde viral vector delivery and in vivo CRISPR/Cas9-mediated somatic genome editing. Genes Dev, 29(14), 1576-1585. doi:10.1101/gad.264861.115

Long Name

CRISPR-associated Protein 9

Alternate Names

Cas9, CRISPR-associated endonuclease Cas9/Csn1, CRISPR-Cas9/Csn1, CRISPR/Cas9, csn1, SPy_1046, SPy1046, SpyCas9

Additional CRISPR-Cas9 Products

Product Documents for CRISPR-Cas9 Antibody (7A9-3A3) [DyLight 650]

Certificate of Analysis

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Product Specific Notices for CRISPR-Cas9 Antibody (7A9-3A3) [DyLight 650]



DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

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