CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - BSA Free
Novus Biologicals, part of Bio-Techne | Catalog # NBP2-36440
Conjugate
Catalog #
Forumulation
Catalog #
Key Product Details
Species Reactivity
Validated:
Bacteria
Cited:
Human, Mouse, Bacteria, Bacterial, Fungi
Applications
Validated:
Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry Whole-Mount, Immunohistochemistry-Frozen, Immunoprecipitation, Knockout Validated, Simple Western, Western Blot
Cited:
IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry-Frozen, Knockout Validated, Simple Western, Western Blot
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG1 kappa Clone # 7A9-3A3
Format
BSA Free
Concentration
1.0 mg/ml
Product Specifications
Immunogen
This CRISPR-Cas9 antibody (7A9-3A3) - N-Terminus was raised against Recombinant Cas9 within the N-terminal region of Streptococcus pyogene. .
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
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.
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 - BSA Free
Simple Western: CRISPR-Cas9 Antibody (7A9-3A3)N-TerminusBSA Free [NBP2-36440]
Simple Western: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - Image shows a specific band for Cas9 (observed molecular weight ~158 kDa) in HeLa Cas9 lysate but not in Hela WT lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.Immunocytochemistry/ Immunofluorescence: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - BSA Free [NBP2-36440]
Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - 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.Immunoprecipitation: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - BSA Free [NBP2-36440]
Immunoprecipitation: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - HEK293T expressing N-terminally Flag-tagged S.pyogenes Cas9 were lysed 72h post transfection by resuspending the cells in Hunt buffer and subjecting to 3 freeze-thaw cycles in liquid nitrogen/ice. Proteins were immunoprecipitated from 100ug of whole cell lysate for 1h at 4C with Cas9 supernatant followed by incubation for 1h at 4C with a 1:1 mixture of protein A/G sepharose beads, or for 2h at 4C with Cas9 ab crosslinked to a 1:1 mixture of protein A/G sepharose beads. Beads were washed 2x with Hunt buffer and 1x with TBS. Bound proteins were eluted by boiling in Laemmli, separated by SDS-PAGE and transferred to nitrocellulose. Membrane was blocked, incubated with Cas9 ab, incubated with HRP anti-mouse secondary. *IgG heavy chainApplications for CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - BSA Free
Application
Recommended Usage
Immunoblotting
reported in scientific literature (PMID 31959836)
Immunocytochemistry/ Immunofluorescence
1:500
Immunohistochemistry-Frozen
reported in scientific literature (PMID 28153089)
Knockout Validated
reported in scientific literature (PMID 31959836)
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
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 8 reviews rated 4.6 using NBP2-36440 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein G purified
Formulation
PBS
Format
BSA Free
Preservative
0.02% Sodium Azide
Concentration
1.0 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
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 - BSA Free
Product Specific Notices for CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - 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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