CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-80679

Novus Biologicals, part of Bio-Techne

Key Product Details

Species Reactivity

Mouse, Bacteria, Bovine, Chicken, Fungi

Applications

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

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 7A9-3A3

Format

Azide and BSA Free

Concentration

1 mg/ml

View all CRISPR-Cas9 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. .

Reactivity Notes

Mouse reactivity reported in multiple pieces of scientific literature.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Theoretical MW

158.4 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free

Western Blot: CRISPR-Cas9 Antibody (7A9-3A3)N-TerminusAzide and BSA Free [NBP2-80679]

Western Blot: CRISPR-Cas9 Antibody (7A9-3A3)N-TerminusAzide and BSA Free [NBP2-80679]

Western Blot: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679] - Analysis of lysate from Cas9 transfected HEK-293T cells using Cas9 antibody clone 7A9-3A3 at 2ug/ml concentration. The signal was developed using HRP-labelled anti-mouse secondary antibody and ECL based detection. Observed molecular weight is ~158 kDa. Image from the standard format of this antibody.
Immunocytochemistry/ Immunofluorescence: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679]

Immunocytochemistry/ Immunofluorescence: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679]

Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679] - Hela cells were transiently transfected with an N-terminally Flag-tagged S. pyogenes Cas9 expression vector. The cells were stained with the Cas9 antibody followed by anti mouse-AF488 coupled secondary antibody. Nuclei were counter-stained with Hoechst 33342. Image from the standard format of this antibody.
Immunohistochemistry-Frozen: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679]

Immunohistochemistry-Frozen: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679]

Immunohistochemistry-Frozen: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679] - Analysis of a formalin fixed 20um thick frozen section of mouse brain with GBM xenograft tumor areas (GBM cells over expressing SpyCas9 through lentivirus infection). CRISPR-Cas9 antibody (clone 7A9-3A3) was used at 1:50 dilution. The signal was detected
View 4 more images

Applications for CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:500

Immunohistochemistry-Frozen

reported in scientific literature (PMID 28153089)

Western Blot

1:1000
Application Notes
IF and IHC use of CRISPR-Cas9 antibody (clone 7A9-3A3) on 4% formaldehyde fixed and 20um thick frozen-/cryo-sections reported in scientific literature (PMID: 28153089)

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

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) - N-Terminus - Azide and BSA Free

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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Product Specific Notices for CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free

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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