RecombinantRabMAb

Recombinant Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19620] to CRISPR-Cas9 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt

Overview

  • Product name

    Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free
    See all CRISPR-Cas9 primary antibodies

  • Description

    Rabbit monoclonal [EPR19620] to CRISPR-Cas9 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Neisseria meningitidis

  • Immunogen

    Recombinant fragment within CRISPR-Cas9 aa 1-300. The exact sequence is proprietary. (Neisseria meningitidis serogroup A/ serotype 4A (strain Z2491)).
    Database link: A1IQ68

  • Positive control

    • ICC/IF: 293T cells transfected with CRISPR-Cas9 (A1IQ68, Neisseria meningitidis serogroup A / serotype 4A (strain Z2491)) with Myc-His tag.

  • General notes

    Ab232503 is the carrier-free version of ab202638. 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.

    ab232503 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    pH: 7.2
    Constituent: PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR19620

  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab232503 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 150 kDa (predicted molecular weight: 124 kDa).
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.
ICC/IF Use at an assay dependent concentration.
Flow Cyt 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.

  • Alternative names

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

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)
    Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)

    Immunocytochemical analysis of agarose-embedded section of 293T (Human epithelial cell line from embryonic kidney) cells transfected with Neisseria meningitidis serogroup A Cas9 (pcDNA3.1(+)-Myc-His) labeling CRISPR-Cas9 with ab202638 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Positive staining on the 293T cells transfected with Neisseria meningitidis serogroup A Cas9 is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is 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 (ab202638).

  • Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)
    Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)

    Immunocytochemical analysis of agarose-embedded sections of 293T (Human epithelial cell line from embryonic kidney) cells transfected with blank pcDNA3.1(+)-Myc-His vector labeling CRISPR-Cas9 with ab202638 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Negative on the 293T cells transfected with blank pcDNA3.1(+)-Myc-His 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 (ab202638).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CRISPR-Cas9 with ab202638 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. 

    No staining on Human tonsil 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 (ab202638).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)

    Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling CRISPR-Cas9 with ab202638 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 (ab202638).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)

    Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling CRISPR-Cas9 with ab202638 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. 

    No staining on rat spleen 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 (ab202638).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)
    Flow Cytometry - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293 (Human epithelial cell line from embryonic kidney) cells transfected with CRISPR-Cas9 (A1IQ68, Neisseria meningitidis serogroup A / serotype 4A (strain Z2491)) with Myc-His tag labeling CRISPR-Cas9 with ab202638 at 1/80 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A]- Isotype control (ab172730) (black). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab202638).

  • Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)
    Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)

    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 (A1IQ68, Neisseria meningitidis serogroup A / serotype 4A (strain Z2491)) with Myc-His tag or Empty vector, labeling CRISPR-Cas9 with ab202638 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    Confocal image showing positive staining on 293T cells transfected with CRISPR-Cas9 (A1IQ68, Neisseria meningitidis serogroup A / serotype 4A (strain Z2491)) with Myc-His tag.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab202638 at 1/100 dilution followed by ab150120  at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by 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 (ab202638).

  • Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)
    Anti-CRISPR-Cas9 antibody [EPR19620] - BSA and Azide free (ab232503)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab232503? Please let us know so that we can cite the reference in this datasheet.

ab232503 has not yet been referenced specifically in any publications.

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