As listed in the 'DESCRIPTION' section under 'Physical Form', this product is supplied as a solution containing 0.01 M phosphate buffered saline, pH 7.4, with 15 mM sodium azide as a preservative. There is no glycerol present.
SAB4200701
Anti-CRISPR/Cas9 antibody, Mouse monoclonal
clone 7A9-3A3, purified from hybridoma cell culture
Synonym(s):
CRISPR-associated protein-9 nuclease, Crisper, Crispr RNA
Skip To
biological source
mouse
Quality Level
antibody form
purified from hybridoma cell culture
antibody product type
primary antibodies
clone
7A9-3A3, monoclonal
form
buffered aqueous solution
packaging
antibody small pack of 25 μL
concentration
~1 mg/mL
technique(s)
immunoblotting: 1-2 μg/mL using whole extracts of human HEK-293T cells over-expressing CAS9 protein
immunofluorescence: 1-2 μg/mL using human HEK-293T cells over-expressing CAS9 protein.
immunoprecipitation (IP): 5-10 μg using whole extract of human HEK-293T cells over-expressing CAS9 protein
isotype
IgG1
shipped in
dry ice
storage temp.
−20°C
target post-translational modification
unmodified
1 of 4
This Item | SAB4200751 | SAB4200756 | SAB4200777 |
|---|---|---|---|
| clone 7A9-3A3, monoclonal | clone 10C11-A12, monoclonal | clone AsCpf-11, monoclonal | clone LbCpf1, monoclonal |
| antibody form purified from hybridoma cell culture | antibody form purified from hybridoma cell culture | antibody form purified from hybridoma cell culture | antibody form purified from hybridoma cell culture |
| biological source mouse | biological source - | biological source mouse | biological source mouse |
| form buffered aqueous solution | form buffered aqueous solution | form buffered aqueous solution | form buffered aqueous solution |
| isotype IgG1 | isotype - | isotype IgG2a | isotype IgG1 |
| technique(s) immunoblotting: 1-2 μg/mL using whole extracts of human HEK-293T cells over-expressing CAS9 protein, immunoprecipitation (IP): 5-10 μg using whole extract of human HEK-293T cells over-expressing CAS9 protein, immunofluorescence: 1-2 μg/mL using human HEK-293T cells over-expressing CAS9 protein. | technique(s) immunoblotting: 1-2 μg/mL using whole extracts of human HEK-293T cells over-expressing CAS9 protein, immunofluorescence: 1.25-2.5 μg/mL using human HEK-293T cells over-expressing CAS9 protein, immunoprecipitation (IP): 5-10 μg/test using whole extract of human HEK-293T cells over-expressing CAS9 protein | technique(s) immunoblotting: 1.25-2.5 μg/mL using purified recombinant AsCpf1 produced in E. coli, immunofluorescence: 1.25-2.5 μg/mL using human HEK-293T cells over-expressing AsCpf1 protein, immunoprecipitation (IP): 2.5-5 μg/test using lysate of human HEK-293T cells over-expressing AsCpf1 protein | technique(s) immunoblotting: 0.6-1.2 μg/mL using human HEK-293T cells over-expressing LbCpf1 protein, immunofluorescence: 0.25-0.5 μg/mL using human HEK-293T cells over-expressing LbCpf1 protein, immunoprecipitation (IP): 1-2.5 μg/test using lysate of human HEK-293T cells over-expressing LbCpf1 protein |
General description
CAS9, also known as clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein-9 nuclease , cas5 and csn1, is the signature gene of the type II CRISPR -RuvC (RNase H-like fold) cas system. CAS9 contains 1388 amino acids. This protein is predicted to contain a RuvC/ ribonuclease (RNase) H domain involved in crRNA maturation and McrA/HNH signature domain involved in the DNA degradation step.
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) belongs to the type II CRISPR/CAS9 system. It is part of an adaptive immune system of the Streptococcus pyogenes SF370, protecting from pathogens targets genes by cleaving the foreign DNA in a sequence-dependent manner.2 The type II CRISPR/Cas system which has been adapted to expression in eukaryotic cells, consists of four genes including the Cas9 (CRISPR-associated proteins) nuclease, two noncoding CRISPR RNAs (crRNAs, or gRNA), trans-activating crRNA (tracrRNA) and a precursor crRNA (pre-crRNA) array. The pre-crRNA contains nuclease guide sequences (spacers) interspaced by identical direct repeats (DRs).
The Cas9 endonuclease can be engineered with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB. In comparison to other genome-editing technologies such as designer zinc fingers (ZFs), transcription activator–like effectors (TALEs) and homing meganucleases, the CRISPR/CAS9 system is a scalable, affordable and easy to engineer. Therefore, the anti-CRISPR/CAS9 antibody can be a useful tool for detecting CRISPR/CAS9 positively transfected cells, reveling DSB sites in the genome and in ChIP (Chromatin Immunoprecipitation) related assays.
The Cas9 endonuclease can be engineered with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB. In comparison to other genome-editing technologies such as designer zinc fingers (ZFs), transcription activator–like effectors (TALEs) and homing meganucleases, the CRISPR/CAS9 system is a scalable, affordable and easy to engineer. Therefore, the anti-CRISPR/CAS9 antibody can be a useful tool for detecting CRISPR/CAS9 positively transfected cells, reveling DSB sites in the genome and in ChIP (Chromatin Immunoprecipitation) related assays.
Immunogen
a recombinant protein within the N-terminal region of Streptococcus pyogene Cas9
Application
Monoclonal Anti-CRISPR/CAS9 recognizes over-expression systems containing CAS9 construct. The antibody successfully recognizes mutant Cas9 variants, including nickase Cas9 and dead Cas9 (dCas9). The antibody may be used in various immunochemical techniques including Immunoblotting, Immunofluorescence and Immunoprecipitation. Monoclonal Anti-CRISPR/CAS9 does not cross react with FnCas9 from Francisella novicida bacteria and Cpf1 proteins from Acidaminococcus sp. (strain BV3L6) and Lachnospiraceae bacterium ND2006.
Biochem/physiol Actions
CAS9 plays a vital role in plasmid DNA interference. It is the only cas protein needed to deliver resistance against foreign DNA. CAS9 stimulates both RNA-guided genome editing and gene regulation in various organisms, but it can facilitate only one activity at a time within any given cell.
Physical form
Supplied as a solution in 0.01 M phosphate buffered saline pH 7.4, containing 15 mM sodium azide as a preservative.
Preparation Note
For continuous use, store at 2-8°C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours.
Other Notes
This product is for R&D use only, not for drug, household, or other uses.
In order to obtain best results in different techniques and preparations we recommend determining optimal working concentration by titration test.
In order to obtain best results in different techniques and preparations we recommend determining optimal working concentration by titration test.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Not finding the right product?
Try our Product Selector Tool.
Storage Class
12 - Non Combustible Liquids
wgk_germany
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
Choose from one of the most recent versions:
Already Own This Product?
Find documentation for the products that you have recently purchased in the Document Library.
Reem Alkhayer et al.
STAR protocols, 5(2), 103045-103045 (2024-05-01)
The unbiased identification of less-abundant transcription factors, which direct the expression of a target gene, is technically challenging. Here, we present a protocol to analyze the locus-specific chromatin-regulating proteome using in situ capture of chromatin interactions by an inactive Cas9
Yen-Ting Liu et al.
Molecular and cellular biology, 43(3), 115-129 (2023-03-22)
CDKN2A/B deletion or silencing is common across human cancer, reinforcing the general importance of bypassing its tumor suppression in cancer formation or progression. In rhabdomyosarcoma (RMS) and neuroblastoma, two common childhood cancers, the three CDKN2A/B transcripts are independently expressed to
Yunjian Wu et al.
Molecular oncology, 17(3), 469-486 (2023-01-07)
Reciprocal interactions between prostate cancer cells and carcinoma-associated fibroblasts (CAFs) mediate cancer development and progression; however, our understanding of the signalling pathways mediating these cellular interactions remains incomplete. To address this, we defined secretome changes upon co-culture of prostate epithelial
L G V Fernandes et al.
Scientific reports, 9(1), 1839-1839 (2019-02-14)
Leptospirosis is a worldwide zoonosis caused by pathogenic bacteria of the genus Leptospira, which also includes free-living saprophyte strains. Many aspects of leptospiral basic biology and virulence mechanisms remain unexplored mainly due to the lack of effective genetic tools available
Jana Ordon et al.
Plant methods, 19(1), 30-30 (2023-03-29)
In plant genome editing, RNA-guided nucleases such as Cas9 from Streptococcus pyogenes (SpCas9) predominantly induce small insertions or deletions at target sites. This can be used for inactivation of protein-coding genes by frame shift mutations. However, in some cases, it
Related Content
Instructions
-
What is the buffered aqueous solution composed of? Just making sure there is no glycerol. Thankyou
1 answer-
Helpful?
-
Active Filters
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
Contact Technical Service