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Antibodies

Cas9 antibody (mAb)

Clone: 7A9-3A3
Aliases: CRISPR/Cas9
Catalog No: 61577 Format: 100 µg $410 Buy Now
Catalog No: 61578 Format: 10 µg $95 Buy Now

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Antibody Type:Monoclonal
Isotype:IgG1k
Purification:Protein G Chromatography
Host:Mouse
Molecular Weight:160 kDa

Applications

Western Blot Validated Immunoprecipitation Validated Immunofluorescence Validated Immunocytochemistry Validated

for Cas9 antibody (mAb) (Clone 7A9-3A3)Application Notes


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

for Cas9 antibody (mAb) (Clone 7A9-3A3)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.

for Cas9 antibody (mAb) (Clone 7A9-3A3)Buffer

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

Also available as AbFlex™ engineered recombinant antibodies.  For details on the corresponding AbFlex Recombinant Antibody, see Catalog No. 91123.

for Cas9 antibody (mAb) (Clone 7A9-3A3)References

 
Cas9 antibody (mAb) tested by immunoprecipitation.

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 immunofluorescence.

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.

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.

for Cas9 antibody (mAb) (Clone 7A9-3A3)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.

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use Cas9 antibody (mAb).

Compatible Secondary Antibodies

Active Motif has developed a variety of high-quality secondary antibodies for most applications, with anti-rabbit and anti-mouse conjugates to a broad line of high-quality fluorescent molecules, as well as horse radish peroxidase (HRP). Visit our Secondary Antibody Conjugates page to find the perfect secondaries for immunofluorescence experiments with Cas9 antibody (mAb).

for Cas9 antibody (mAb) (Clone 7A9-3A3)Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Upon receipt, unconjugated antibodies may be stored at -20°C for up to 2 years. Fluorophore- & enzyme-conjugated antibodies should be stored at 4°C. Fluorophore-conjugated antibodies should be protected from light. Keep reagents on ice when not in storage; to avoid repeated freeze/thaw cycles, we recommend aliquoting items that will be stored frozen into single-use fractions prior to freezing.

for Cas9 antibody (mAb) (Clone 7A9-3A3)Guarantee

This product is guaranteed for 6 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;
  • 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;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting
 

Technical Data Sheet

Cas9 antibody (mAb)
 

Data Thumbnails

Cas9 antibody (mAb) tested by immunoprecipitation.

Immunoprecipitation of Cas9 mAb.
(Click image to enlarge and see details.)

Cas9 antibody (mAb) tested by immunofluorescence.

Immunofluorescence stain of Cas9 mAb.
(Click image to enlarge and see details.)

Cas9 antibody (mAb) tested by Western blot.

Western blot of Cas9 mAb.
(Click image to enlarge and see details.)