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ABC1490 Anti-Cas9 Antibody, D10A/H840A Mutant

Detect Cas9 D10A/H840A nickase mutant using this rabbit polyclonal Anti-Cas9, D10A/H840A Mutant, Cat. No. ABC1490, validated for use in Immunoprecipitation and Western Blotting.

Anti-Cas9 Antibody, D10A/H840A Mutant MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Research Category: Secondary & Control Antibodies Gene Symbol: cas9, csn1 UniProt Number: Q99ZW2
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ABC1490
100 μg  
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      Overview

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      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      AIP, WBRbAffinity PurifiedPolyclonal Antibody
      Description
      Catalogue NumberABC1490
      DescriptionAnti-Cas9 Antibody, D10A/H840A Mutant
      Alternate Names
      • CRISPR-associated endonuclease Cas9/Csn1
      • D10A/H840A mutant
      • SpyCas9
      • D10A/H840A mutant
      Background InformationCRISPR-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. Mutations of the catalytic residues (D10A in RuvC and H840A in HNH) convert Cas9 into DNA nickases (Cas9n) that cleave only the strand complementary to the crRNA, minimizing off-target activity.
      References
      Product Information
      FormatAffinity Purified
      PresentationPurified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
      Quality LevelMQ100
      Applications
      ApplicationDetect Cas9 D10A/H840A nickase mutant using this rabbit polyclonal Anti-Cas9, D10A/H840A Mutant, Cat. No. ABC1490, validated for use in Immunoprecipitation and Western Blotting.
      Key Applications
      • Immunoprecipitation
      • Western Blotting
      Application NotesImmunoprecipitation Analysis: 10 µg from a representative lot immunoprecipitated exogenously expressed Cas9 nickase D10A mutant (with C-terminal Myc-DDK tag) from 500 µg lysate of transfected HEK293 cells.
      Biological Information
      ImmunogenA mixture of two KLH-conjugated peptides corresponding to the Cas9 target sites sequences with D10A and H840A mutations.
      ConcentrationPlease refer to lot specific datasheet.
      HostRabbit
      SpecificityThis polyclonal antibody will detect Cas9 with D10A mutation in the RuvC-I region and/or H840A mutation in the HNH domain (e.g. AKA60242.1), but not Cas9 with neither D10A nor H840A mutation (e.g. NP_269215).
      Species Reactivity
      • All
      Species Reactivity NoteStreptococcus pyogenes.
      Antibody TypePolyclonal Antibody
      Entrez Gene Number
      Gene Symbol
      • cas9
      • csn1
      Purification MethodAffinity purified.
      UniProt Number
      Molecular Weight~160 kDa observed. 158.3 kDa calculated (based on AKA60242.1). Uncharacterized bands may be observed in some lysate(s).
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceEvaluated by Western Blotting in lysate from HEK293 transfectants expressing Cas9 nickase D10A.

      Western Blotting Analysis: 2 µg/mL of this antibody detected exogenously expressed Cas9 nickase D10A mutant (with C-terminal Myc-DDK tag) in 10 µg of lysate from transfected HEK293 cells.
      Usage Statement
      • 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.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
      Packaging Information
      Material Size100 μg
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      ABC1490 04054839108761