Recombinant
RabMAb

Recombinant Anti-CRISPR-Cas9 antibody [EPR19795] - BSA and Azide free (ab223153)

Overview

  • Product name

    Anti-CRISPR-Cas9 antibody [EPR19795] - BSA and Azide free
    See all CRISPR-Cas9 primary antibodies
  • Description

    Rabbit monoclonal [EPR19795] to CRISPR-Cas9 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Staphylococcus aureus
  • Immunogen

    Recombinant fragment aa 1-400. The exact sequence is proprietary. (subsp. aureus).
    Database link: J7RUA5

  • Positive control

    • WB: HEK-293T whole cell lysate transfected with Myc-His tagged CRISPR-Cas9 (J7RUA5, Staphylococcus aureus) vector. ICC/IF: HEK-293T cells transfected with GFP-tagged CRISPR-Cas9 (J7RUA5, Staphylococcus aureus) expression construct. Flow Cyt: HEK-293T cells transfected with GFP-tagged CRISPR-Cas9 (J7RUA5, Staphylococcus aureus) expression construct. IP: HEK-293T cell lysate transfected with GFP-tagged CRISPR-Cas9 (J7RUA5, Staphylococcus aureus) vector.
  • General notes

    Ab223153 is the carrier-free version of ab203943. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab223153 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19795
  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab223153 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 Use at an assay dependent concentration. Detects a band of approximately 124 kDa (predicted molecular weight: 124 kDa).
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.

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

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cell line from embryonic kidney) cells labeling CRISPR-Cas9 with ab203943 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing positive staining on HEK-293T cells transfected with a GFP-tagged CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) expression construct.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203943).

    • Flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293T (Human epithelial cell line from embryonic kidney) cells transfected with a GFP tagged CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) vector, labeling CRISPR-Cas9 with ab203943 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black). Goat anti rabbit IgG (Alexa Fluor® 647) at 1/2000 dilution was used as the secondary antibody.

      Note: The image is obtained by gating the GFP-tag positive population.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203943).

    • CRISPR-Cas9 was immunoprecipitated from 0.35 mg of HEK-293T (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with a GFP tagged CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) vector,  with ab203943 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab203943 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: HEK-293T whole cell lysate transfected with a GFP tagged CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) vector, 10 μg (Input).

      Lane 2: ab203943 IP in HEK-293T whole cell lysate transfected with a GFP tagged CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) vector.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab203943 in HEK-293T whole cell lysate transfected with a GFP tagged CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) vector.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 10 seconds.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203943).

    References

    ab223153 has not yet been referenced specifically in any publications.

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