Immunogen:

E. coli-derived Cas9
Asp2-Asp1368
Accession # Q99ZW2

Applications:

Western Blot

Label:

Alexa Fluor 488 (Excitation=488 nm, Emission=515-545 nm)

Specificity:

Detects Cas9 in direct ELISAs and Western blots.

View Full Technical Details for CRISPR-Cas9 Alexa Fluor® 488-conjugated Antibody

Technical Data
Citations / Reviews

Applications

	Recommended Concentration	Sample
Western Blot	Optimal dilution of this antibody should be experimentally determined.

Please Note: Optimal dilutions should be determined by each laboratory for each application.

Formulation, Preparation and Storage

Purification	Protein A or G purified
Formulation	Supplied 0.2mg/ml in 1X PBS with RDF1 and 0.09% Sodium Azide
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Protect from light. Do not freeze. 12 months from date of receipt, 2 to 8 °C as supplied

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Background: CRISPR-Cas9

Streptococcus pyogenes Cas9 (CRISPR associated protein 9) is a 160 kDa RNA guided endonuclease that introduces site specific cleavage of double strand DNA (1). It is part of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) system found in many bacteria such as S. pyogenes and most archaea, which provide adaptive immunity against invading mobile genetic elements (such as viruses, transposable elements and conjugative plasmids) (2, 3). Upon viral infection, short viral DNA (known as "spacers") integrate into the host genome between CRISPR repeats, and RNA sequences (guide RNA or gRNA) with this genetic information help guide Cas9 protein to recognize and cut foreign DNA. Cas9 protein undergoes conformational changes upon gRNA binding that shift from non-DNA binding conformation into an active DNA binding conformation. In the Cas9-gRNA complex, the gRNA sequence remains accessible to interact with free DNA, and the extent to which the gRNA spacer and target DNA segment (known as "protospacer") match will determine the cut site (4). The presence of a 5′-NGG-3′ protospacer adjacent motif (PAM) sequence immediately downstream of protospacers is required for Cas9 cleavage of the foreign DNA. PAM is absent in bacterial CRISPR loci, therefore preventing cleavage of the host genome (4). Cas9 associates with other proteins of the acquisition machinery (Cas1, Cas2 and Csn2), presumably to provide PAM specificity to this process (5). This RNA guided nuclease system called CRISPR/Cas (CRISPR associated protein) has been widely applied to genome engineering with increased efficiency (6). The attached nuclear localization signals(NLSs) on the chimeric protein ensures nuclear compartmentalization in mammalian cells during gene editing (7).

Item	Value
Long Name	CRISPR-associated Protein 9
Entrez Gene IDs	901176 (S. pyogenes)
Alternate Names	CRISPR, CRISPR-Cas9, CRISPR-Cas9/Csn1, CRISPR-associated endonuclease Cas9/Csn1, CRISPR-associated protein 9 nuclease, CRISPR/Cas9, Cas9, SPy1046, SPy_1046, SpyCas9, csn1

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CRISPR-Cas9 Alexa Fluor® 488-conjugated Antibody

R&D Systems, part of Bio-Techne | Catalog # FAB10252G

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