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Your Position: Home > Enzyme > CAS9 > CA9-S5149

GENPower™ NLS-Cas9 Nuclease

  • Synonym
    CAS9
  • Source
    GENPower™ NLS-Cas9 Nuclease (CA9-S5149) is expressed from E.coli cells for genome editing applications with CRISPR technology with N-terminus and C-terminus NLS for improved transport to the nucleus . It is ideal for in vitro and in vivo reactions. It contains AA Asp 2 - Asp 1368 (Accession # Q99ZW2-1).
    Predicted N-terminus: Met 1
  • Molecular Characterization
    CAS9 Structure

    This protein carries a polyhistidine tag at the N-terminus

    The protein has a calculated MW of 164.8 Kda. The protein migrates as 140-150 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.

  • Endotoxin
    Less than 0.01 EU per μg by the LAL method.
  • Sterility
    The sterility testing was performed by membrane filtration method.
  • Purity

    >95% as determined by SDS-PAGE.

    >95% as determined by SEC-MALS.

  • Formulation

    Supplied as 0.2 μm filtered solution in 20 mM Tris, 300 mM NaCl, 1 mM EDTA, 1 mM TCEP with Glycerol as protectant.

    Contact us for customized product form or formulation.

  • Shipping

    This product is supplied and shipped as sterile liquid solution with dry ice, please inquire the shipping cost.

  • Storage

    Please avoid repeated freeze-thaw cycles.

    This product is stable after storage at:

    1. The product MUST be stored at -20°C or lower upon receipt;
    2. -20°C for 3 months under sterile conditions.
SDS-PAGE
CAS9 SDS-PAGE

GENPower™ NLS-Cas9 Nuclease on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.

SEC-MALS
CAS9 MALS images

The purity of GENPower™ NLS-Cas9 Nuclease(Cat. No. CA9-S5149) is more than 95% and the molecular weight of this protein is around 150-180 kDa verified by SEC-MALS.

Bioactivity

GENPower™ NLS-Cas9 Nuclease is evaluated in an in vitro DNA cleavage assay on a DNA fragment containing the target sequence. The activity of the GENPower™ NLS-Cas9 Nuclease is greater than 90%.

  • Background
    CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids),CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. 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 this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA; Cas9 only stabilizes the pre-crRNA:tracrRNA interaction and has no catalytic function in RNA processing. Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). The target strand not complementary to crRNA is first cut endonucleolytically, then trimmed 3-5 exonucleolytically. DNA-binding requires protein and both gRNAs, as does nuclease activity. Cas9 recognizes the protospacer adjacent motif (PAM) in the CRISPR repeat sequences to help distinguish self versus nonself, as targets within the bacterial CRISPR locus do not have PAMs. DNA strand separation and heteroduplex formation starts at PAM sites; PAM recognition is required for catalytic activity.
  • Clinical and Translational Updates
  • Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.

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