Recombinant
RabMAb

Recombinant Anti-CRISPR-Cas9 antibody [EPR18991] - BSA and Azide free (ab232379)

Overview

  • Product name

    Anti-CRISPR-Cas9 antibody [EPR18991] - BSA and Azide free
    See all CRISPR-Cas9 primary antibodies
  • Description

    Rabbit monoclonal [EPR18991] to CRISPR-Cas9 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Streptococcus pyogenes
  • Immunogen

    Recombinant fragment within Streptococcus pyogenes CRISPR-Cas9 aa 800-1000. The exact sequence is proprietary. (Serotype M1).
    Database link: Q99ZW2

  • Positive control

    • ICC/IF: 293T cells transfected with CRISPR-Cas9 with GFP-tag.
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®). 

    ab232379 is a PBS-only buffer format of ab189380. Please refer to ab189380 for recommended dilutions, protocols, and image data.

    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.

    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

    EPR18991
  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab232379 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 184 kDa (predicted molecular weight: 158 kDa).
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    • Flow cytometry analysis of HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate (transfected with CRISPR-Cas9 (Q99ZW2, Streptococcus pyogenes serotype M1) with GFP-Myc tag) labelling CRISPR-Cas9 (red) with ab189380 at dilution of 1/70. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde. Isotype control antibody was (ab172730) Rabbit monoclonal IgG (black).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).

    • Immunocytochemical analysis of agarose-embedded sections of 293T (Human epithelial cell line from embryonic kidney) cells transfected with Streptococcus pyogenes serotype M1 Cas9 (pcDNA3.1(+)-GFP-Myc) labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Positive staining on 293T cells transfected with Streptococcus pyogenes serotype M1 Cas9 (pcDNA3.1(+)-GFP-Myc) is observed.

      Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).

    • Immunocytochemical analysis of agarose-embedded sections of 293T (Human epithelial cell line from embryonic kidney) cells transfected with blank pcDNA3.1(+)-GFP-Myc vector labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Negative on 293T cells transfected with blank pcDNA3.1(+)-GFP-Myc vector.

      Counter stained with Hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).

    • Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. 

      No staining on Human stomach is observed.

      Counter stained with Hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).

    • Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. 

      No staining on mouse stomach is observed.

      Counter stained with Hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).

    • Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. 

      No staining on rat cerebrum is observed.

      Counter stained with Hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T (Human epithelial cell line from embryonic kidney) cells transfected with CRISPR-Cas9-GFP or GFP only, labeling CRISPR-Cas9 with ab189380 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 647) (ab150079) secondary antibody at 1/1000 dilution.

      Confocal image showing positive staining on 293T cells transfected with CRISPR-Cas9 with GFP-tag.

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

      The negative controls are as follows:-

      -ve control 1: ab189380 at 1/500 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.

      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150079 (Alexa Fluor® 647 Goat Anti-Rabbit IgG H&L) 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 (ab189380).

    References

    ab232379 has not yet been referenced specifically in any publications.

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