Anti-CRISPR-Cas9 antibody [7A9-3A3] (HRP) (ab202580)

Overview

  • Product name

    Anti-CRISPR-Cas9 antibody [7A9-3A3] (HRP)
    See all CRISPR-Cas9 primary antibodies
  • Description

    Mouse monoclonal [7A9-3A3] to CRISPR-Cas9 (HRP)
  • Host species

    Mouse
  • Conjugation

    HRP
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Streptococcus pyogenes
  • Immunogen

    Recombinant fragment corresponding to Streptococcus pyogenes CRISPR-Cas9 (N terminal).

  • Positive control

    • WB: S2 cells transfected with CRISPR-Cas9

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. Store In the Dark.
  • Storage buffer

    pH: 7.40
    Preservative: 0.1% Proclin
    Constituents: PBS, 1% BSA, 30% Glycerol
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    7A9-3A3
  • Isotype

    IgG1
  • Light chain type

    kappa

Applications

Our Abpromise guarantee covers the use of ab202580 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/5000. Detects a band of approximately 180 kDa (predicted molecular weight: 160 kDa).

We recommend using 3% milk as the blocking agent for Western blot.

Target

  • Relevance

    [FUNCTION] 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) (Probable). 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. Subsequently 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, then trimmed by 3'-5' exonucleolytically. DNA-binding requires protein and both RNA species. Cas9 probably recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus nonself.
  • Database links

    • Alternative names

      • Cas9 antibody
      • CRISPR-associated endonuclease Cas9/Csn1 antibody
      • CRISPR-Cas9/Csn1 antibody
      • csn1 antibody
      • SpyCas9 antibody
      see all

    Images

    • All lanes : Anti-CRISPR-Cas9 antibody [7A9-3A3] (HRP) (ab202580) at 1/5000 dilution

      Lane 1 : Cas 9 negative control (S2 non-transfected cell lysate)
      Lane 2 : Cas 9 transfected cells (S2 cells transfected with CRISPR-Cas9 lysate)

      Lysates/proteins at 20 µg per lane.

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 160 kDa
      Observed band size: 180 kDa
      why is the actual band size different from the predicted?


      Exposure time: 2 minutes


      This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab202580 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

    References

    This product has been referenced in:

    • Koch B  et al. Generation and validation of homozygous fluorescent knock-in cells using CRISPR-Cas9 genome editing. Nat Protoc 13:1465-1487 (2018). Read more (PubMed: 29844520) »
    • Zhang S & Voigt CA Engineered dCas9 with reduced toxicity in bacteria: implications for genetic circuit design. Nucleic Acids Res 46:11115-11125 (2018). Read more (PubMed: 30289463) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris)
    Loading amount
    20 µg
    Treatment
    1 µg/mL Doxacycline (Lane 2)
    Specification
    HeLa
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

    Abcam user community

    Verified customer

    Submitted Nov 19 2018

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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