Recombinant
RabMAb

Recombinant Anti-CRISPR-Cas9 antibody [EPR19619-18] (ab202637)

Overview

  • Product name

    Anti-CRISPR-Cas9 antibody [EPR19619-18]
    See all CRISPR-Cas9 primary antibodies
  • Description

    Rabbit monoclonal [EPR19619-18] to CRISPR-Cas9
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, WBmore details
  • Species reactivity

    Reacts with: Other species
  • Immunogen

    Recombinant fragment within CRISPR-Cas9 aa 150-600. The exact sequence is proprietary. (Streptococcus thermophilus).
    Database link: G3ECR1

  • Positive control

    • WB: HEK-293 whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus, N–terminal) with GFP-tag; HEK-293 whole cell lysate transfected with CRISPR-Cas9 (Q03JI6, Streptococcus thermophilus (strain ATCC BAA-491/LMD-9)) with His-tag; HEK-293 whole cell lysate transfected with CRISPR-Cas9 (A1IQ68, Neisseria meningitidis serogroup A) with His-tag. IP: 293T cells transfected with Streptococcus thermophilus CRISPR-Cas9-myc whole cell lysate.
  • General notes

     

     

    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

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19619-18
  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab202637 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
IP 1/30.
WB 1/5000. Detects a band of approximately 164 kDa (predicted molecular weight: 164 kDa).

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

  • All lanes : Anti-CRISPR-Cas9 antibody [EPR19619-18] (ab202637) at 1/5000 dilution

    Lane 1 : Untransfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate (control)
    Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (Q99ZW2, Streptococcus pyogenes serotype M1) with GFP-tag
    Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus, N–terminal aa1-800) with GFP-tag
    Lane 4 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus, C–terminal aa801-1409) with GFP-tag

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 164 kDa


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The immunogen is located at the N-terminus.

  • All lanes : Anti-CRISPR-Cas9 antibody [EPR19619-18] (ab202637) at 1/5000 dilution

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus, N–terminal) with GFP-tag
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 4 : Rat embryo lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 164 kDa


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-CRISPR-Cas9 antibody [EPR19619-18] (ab202637) at 1/5000 dilution

    Lane 1 : Untransfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate (control)
    Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) transfected with CRISPR-Cas9 (Q03JI6, Streptococcus thermophilus (strain ATCC BAA-491/LMD-9)) with His-tag
    Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (A1IQ68, Neisseria meningitidis serogroup A) with His-tag

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 164 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-2: 5 seconds; Lane 3: 3 minutes.

  • CRISPR-Cas9 was immunoprecipitated from 1mg of 293T (Human epithelial cell line from embryonic kidney) cells transfected with Streptococcus thermophilus CRISPR-Cas9-myc whole cell lysate with ab202637 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab202637 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1:  293T cells transfected with Streptococcus thermophilus CRISPR-Cas9-myc whole cell lysate, 10µg (Input).

    Lane 2: ab202637 IP in 293T cells transfected with Streptococcus thermophilus CRISPR-Cas9-myc whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab202637 in 293T cells transfected with Streptococcus thermophilus CRISPR-Cas9-myc whole cell lysate.

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

    Exposure time: 1 second.

References

ab202637 has not yet been referenced specifically in any publications.

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