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CRISPR/Cas9 Monoclonal Antibody [7A9]

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For antibody-based studies in CRISPR/Cas9 genome editing and genetic engineering

Citations (3) | (1) | Write a Review
Western Blot: Shown are the results of WB on protein extracts from untransfected (A) and transfected (B) HEK293 cells using the Anti-CRISPR-Cas9 mAb.
Immunofluorescence: Hela cells were transiently transfected with an N-terminally Flag-tagged S. pyogenes Cas9 expression vector. The cells were stained with the Anti-Cas9 monoclonal antibody followed by anti mouse-AF488 coupled secondary antibody. Nuclei were counter-stained with Hoechst 33342.
Immunoprecipitation: HEK293T expressing N-terminally Flag-tagged S.pyogenes Cas9 were lysed 72 hours post transfection. Proteins were immunoprecipitated from 100 μg of whole cell lysate for 1H at 4
Application: ELISA, IF, IP, WB
Areas of Research: Other Antibodies
Clonality: Monoclonal
Conjugate: Unconjugated
Host: Mouse
Isotype: IgG1 Kappa
Purification: Protein G Purified
Reactivity: Species Independent
100% Guarantee: 6 months
Catalog No.SizePriceQty
A-9000-01010 µg $59.00 
A-9000-05050 µg $163.00 
A-9000-100100 µg $287.00 
Availability: Usually Ships in 1 Day or Same day delivery Same Day NY Delivery 
Product Overview

Background
The discovery of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) and Cas9 (CRISPR associated system or CRISPR associated protein 9 nuclease) found in bacteria to work as a defense mechanism against foreign DNA has proven to be an invaluable tool to target and modify a genetic sequence in gene editing and genome engineering applications. The system, known as CRISPR/Cas9, allows for sequence-specific cleavage of a targeted genomic locus by delivering the RNA-guided nuclease (Cas9) and appropriate guide RNAs (gRNA) into a cell. In addition, Protospacer Adjacent Motif (PAM) sequence immediately following the specificity sequence is necessary for successful binding of the Cas9 nuclease.

Concentration 
1 mg/ml

Description 
Mouse monoclonal antibody raised against CRISPR/Cas9, clone 7A9, generated with synthesized peptide corresponding to sequence of Streptococcus pyogene (S. pyogenes) CRISPR-associated endonuclease Cas9/Csn1. This Anti-Cas9 mAb can detect CRISPR/Cas9 expression in target cells by WB, IF, IP, or ELISA to confirm and verify whether gRNA and Cas9 vectors are successfully transfected.

Specificity 
Recognizes both Cas9 and dCas9 (nuclease deficient Cas9)

Reactivity
Species Independent

Isotype 
IgG1/Kappa

Immunogen
Recombinant N-terminal fragment of S. pyogenes Cas9 protein expressed in E. coli.

Formulation 
PBS, 30% Glycerol

Storage  
Store at 4°C for short term (1-2 weeks). For long term storage, aliquot and then store at −20°C . Avoid multiple freeze/thaw cycles.

Purity
Protein G Purified

Alternative Names 
Anti-Cas9, Anti-CRISPR, Anti-CRISPR/Cas9, CRISPR antibody, Cas9 antibody, Cas9 7A9, 7A9-3A3

Application
WB (1:500), IP (2 µg/10^6 cells), IF (1:500), ELISA (1:1000-1:2000)

User Guide & MSDS

[User Guide]*
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Customer Reviews

Rating By s***********@dartmouth.edu Verified Purchase Reviewed on: Tuesday 23 June, 2015
Application Description
Western Blot detection of Cas9 protein expression in A. fumigatus Af293 transformed strains.

Procedural Details
For this membrane, I used 1:500 dilution of the Cas9 mAb in blocking buffer (0.1% PBST with 5% non-fat dry milk) and incubated the membrane overnight at 4 C. For my secondary antibody, I used 1:3000 dilution of goat anti-mouse IgG(H+L) HRP-conjugated antibody (Bio-Rad).

Lane 1: protein ladder (on membrane), lane 2: wild-type Af293, lanes 3-5: cas9 transformed Af293 #1-3; short and long exposure, gel loaded with 10 ug protein per well

Other Thoughts
Cas9 protein is the dominant signal when Cas9 is under a constitutive promoter. The 10 s exposure is quite clean, and I overexposed it to get a sense for what other non-specific proteins the Ab might be binding to.


Western blot, 10 second exposure and 3 min over-exposure of same gel
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