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CRISPR-Cas9 (Active) Protein

Spezies: Streptococcus pyogenes Wirt: Escherichia coli (E. coli) Recombinant > 95 % pure as determined by SDS-PAGE with Coomassie Blue detection. GEEN, IVCA, AbP Active
Produktnummer ABIN3071563
  • Target
    CRISPR-Cas9
    Protein-Typ
    Recombinant
    Biologische Aktivität
    Active
    Spezies
    • 6
    • 1
    Streptococcus pyogenes
    Quelle
    • 7
    Escherichia coli (E. coli)
    Applikation
    Genome Editing with Engineered Nucleases (GEEN), In vitro Cleavage Assay (IVCA), Antibody Production (AbP)
    Spezifität
    Activity test
    Cas9 site-specific digestion:
    We used in vitro digestion of a linearized plasmid to determine the activity of the Cas9 nuclease. It is a sensitive assay for GenCrispr Cas9 quality control. The linearized plasmid containing the target site:
    (CATCATTGGAAAACGTTCTT)
    can be digested with gRNA:
    (CAUCAUUGGAAAACGUUCUUGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGCUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUUUU)
    and GenCrispr Cas9. Two cleavage DNA fragments (812 bp and 1898 bp) are determined by agarose gel electrophoresis. A 20 μL reaction in 1xCas9 Nuclease Reaction Buffer containing 160 ng linearized plasmid, 40 nM gRNA and 20 nM GenCrispr Cas9 for 2 hours at 37 °C results in 90 % digestion of linearized plasmid as determined by agarose gel electrophoresis.
    Produktmerkmale
    GenCrispr Cas9 Nuclease is produced by expression in an E. colistrain carrying a plasmid encoding the Cas9 gene from Streptococcus pyogenes without nuclear localization signal (NLS).
    Aufreinigung
    purified
    Reinheit
    > 95 % pure as determined by SDS-PAGE with Coomassie Blue detection.
    Bestandteile
    GenCrispr Cas9 Nuclease
    10X Reaction Buffer
  • Applikationshinweise
    Screening for highly efficient and specific targeting gRNAs by in vitro DNA cleavage using Cas9 Nuclease from S. pyogenes
    Highly purified Cas9 antigen could be used for specific antibody production.
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Konzentration
    0.2 mg/mL
    Buffer
    10X Reaction Buffer: 200 mM HEPES, 1M NaCl, 50 mM MgCl2, 1 mM EDTA, pH 6.5 at 25 °C.
    1X Storage Buffer: 10 mM Tris, 300 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50 % Glycerol pH 7.4 at 25 °C
    Konservierungsmittel
    Dithiothreitol (DTT)
    Vorsichtsmaßnahmen
    This product contains Dithiothreitol (DTT): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Lagerung
    -20 °C
    Informationen zur Lagerung
    GenCrispr Cas9 is supplied with 1X storage buffer (10 mM Tris, 300 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol PH 7.4 at 25°C) and is recommended to be stored at -20°C.br/>Diluent Compatibility: Diluent Buffer: 300 mM NaCl, 10 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, 500 μg/ml BSA and 50% glycerol. (pH 7.4 at 25°C).
  • Target
    CRISPR-Cas9
    Andere Bezeichnung
    Cas9 Nuclease
    Hintergrund
    GenCrispr Cas9 Nuclease is the recombinant Streptococcus pyogenes Cas9 (wt) protein purified from E. coli that can be used for genome editing by inducing site-specific double stranded breaks in double stranded DNA. Cas9 protein forms a very stable ribonucleoprotein (RNP) complex with the guide RNA (gRNA) component of the CRISPR/Cas9 system. The RNP complex recognizes the target site by matching gRNA with the genomic DNA sequence and produces DNA breaks within 3 bases from the NGG PAM (Protospacer Adjacent Motif). With GenCrispr Cas9 nuclease, customers can screen for highly efficient gRNA in vitro using DNA cleavage assays. The high purity Cas9 protein can also be used for antibody production.
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