Cas9, CT (CRISPR-Associated Endonuclease Cas9)

Size

£748.00

SKU:: 046205
Purity:: Purified by Protein G affinity chromatography.
Storage Conditions:: -20°C
Supplier:: United States Biological
Host:: Mouse
Immunogen:: His-tagged recombinant protein corresponding to S.pyogenes Cas9 C-terminal fragment.
Clone:: 10C11-A12
Format:: Supplied as a liquid in 0.1M Tris-Glycine, pH 7.4, 150mM sodium chloride, 0.05% sodium azide.
Extra Details:: CRISPR-associated endonuclease Cas9/Csn1 (UniProt Q99ZW2; also known as SpyCas9) is encoded by the Streptococcus pyogenes serotype M1 gene cas9 (also known as csn1) gene (Gene ID 901176). Clustered regularly interspaced short palindromic repeats (CRISPRs) are DNA loci containing short repetitions of base sequences complementary to antecedent mobile elements and target invading nucleic acids. Each repetition is followed by short segments of spacer DNA from previous exposures to a virus. Cas9 is an essential endonuclease in Streptococcus pyogenes serotype M1's CRISPR immune system that confers resistance to foreign genetic elements such as plasmids and phages. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems, correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc), and Cas9. Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer. The target strand not complementary to crRNA is first cut endonucleolytically, and then trimmed 3'-5' exonucleolytically. Applications: Suitable for use in Western Blot, Immunoprecipitation and Immunocytochemistry. Other applications not tested. Recommended Dilutions: Immunocytochemistry: 33.5ug/ml detected doxycycline-induced expression of exogenously transfected S. pyogenes Cas9 construct under the control of the PTight (Tet-ON) promoter by fluorescent immunocytochemistry staining of methanol-fixed, acetone-permeabilized HeLa transfectants (Courtesy of Dr. Stefan Schuechner and Dr. Egon Ogris, Medical University of Vienna, Austria). Western Blot: 0.5ug/ml detected the exogenously expressed Cas9 in 10ug lysate from transfected HeLa cells. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Shipping Conditions:: Blue Ice