Cas9 antibody (mAb)

RRID: AB_2793684
Clone: 7A9-3A3
Aliases: CRISPR/Cas9
Catalog No: 61577 Format: 100 µg $510 Buy
Catalog No: 61978 Format: 50 µg $295 Buy
Catalog No: 61578 Format: 10 µg $125 Buy

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Antibody Type:
Monoclonal
Isotype:
IgG1, k
Purification:
Protein G Chromatography
Host:
Mouse
Molecular Weight:
160 kDa
Reactivity:
Human, Not Species Specific

Applications

ChIP Validated Western Blot Validated Immunoprecipitation Validated Immunofluorescence Validated Immunocytochemistry Validated

Application Notes

Applications Validated by Active Motif:
IP: 5 µg per IP
ICC/IF: 0.5 - 2 µg/ml dilution
WB: 0.1 - 1 µg/ml dilution

Published Applications

The following applications have been published using this antibody. Unless noted above, Active Motif may not have validated the antibody for use in these applications:

ChIP-qPCR, ChIP-Seq
IHC(F)/IF
FC

View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this antibody.

Immunogen

This antibody was raised against a recombinant protein within the N-terminal region of Streptococcus pyogene Cas9. This antibody should recognize Cas9 and dCas9 based on the antigen design.

Buffer

Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

 

Cas9 antibody (mAb) tested by Immunoprecipitation.
5 µg of Cas9 antibody (mAb) was used to immunoprecipitate Cas9 from 100 µg of whole cell extracts after 72 hours of transient transfection of HEK293 cells with FLAG-Tagged Cas9. The immunoprecipitated protein was detected by Western blotting using Cas9 antibody (mAb) at a 0.5 µg/mL dilution.

Cas9 antibody (mAb) tested by Immunoflourescence.
HeLa cells were transiently transfected with FLAG-Tagged Cas9 for 48 hours and then stained. Top: Cas9 antibody (mAb) at a 0.5 µg/ml dilution. Bottom: Stained with Hoechst.

Cas9 antibody (mAb) tested by Western blot.
HEK293 cells were transiently transfected with Myc-Tagged Cas9 (lane 2) or untransfected (lane 1), lysates prepared 40 hours post transfection, and probed with Cas9 antibody (mAb) at a 0.5 µg/ml dilution.

Background

Cas9 is a nuclease from Streptococcus pyogenes that can be targeted to particular DNA sequences through a guide RNA that results in double-stranded breaks in DNA. Cas9 is part of the CRISPR/Cas9 gene-editing system that can create a DNA break at a specific location with the genome.

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.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with Cas9 antibody (mAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • WB = Western Blotting