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CRISPR/Cas9 Polyclonal Antibody

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For antibody-based studies in gene editing and genome engineering

Citations (2) | (1) | Write a Review
Western blot analysis on Cas9 transfected 293 cells (+) and un-transfected 293 cells (-).
Specific detection of Cas9 nuclease by CRISPR/Cas9 pAb (Cat. #A-1111) via ELISA. Cas9 nuclease was added into the assay wells and detected with CRISPR/Cas9 pAb (at 1:1000 dilution).
Successful immunoprecipitation of Cas9 nuclease by CRISPR/Cas9 pAb (Cat. #A-1111). 200 ng of nuclease was immunoprecipitated with 0.5 ug of CRISPR/Cas9 pAb. Rabbit non-immune IgG was used as control.
Immunofluorescence analysis of CRISPR/Cas9 transfected 293 cells. The cell expressing CRISPR/Cas9 can be detected using rabbit anti-CRISPR/Cas9 (1:500) followed by FITC (1:1000) goat anti-rabbit IgG.
Areas of Research: Other Primary Antibodies
Clonality: Other Antibodies
Conjugate: Unconjugated
Host: Rabbit
Isotype: IgG
100% Guarantee: 6 months
Catalog No.SizePriceQty
A-1111-01010 µg $71.00  Discontinued
A-1111-05050 µg $195.00  Discontinued
A-1111-100100 µg $349.00  Discontinued
Availability: Discontinued 
Product Overview


We recommend our monoclonal Anti-Cas9-CRISPR (Cat #A-9000).


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 
Rabbit polyclonal antibody to CRISPR/Cas9, generated with synthesized peptide corresponding to sequence of Streptococcus pyogene (S. pyogenes) CRISPR-associated endonuclease Cas9/Csn1, with further optimization for IP and ChIP due to recognition of multiple epitopes. This antibody can (a) detect CRISPR/Cas9 expression in target cells by WB, IF, or ELISA to confirm and verify whether gRNA and Cas9 vectors are successfully transfected; or (b) detect genome-wide off- and on-target effects of CRISPR/Cas9 on specific gene sites by Cas9 ChIP-seq in CRISPR genome editing.

Specificity 
Recognizes both Cas9 and dCas9 (nuclease deficient Cas9)

Reactivity:
Species Independent

Isotype 
IgG

Formulation 
Liquid containing 0.05% Sodium Azide, 1X TBS (pH7.4), 0.5% BSA, 40% Glycerol

Storage  
Store at 4°C for 3 weeks. For long term storage, aliquot and then store at −20°C or −80°C. Avoid multiple freeze/thaw cycles.

Purity 
Protein A purified

Handling Recommendations 
For maximum recovery of the products, centrifuge the vial prior to opening the cap

Alternative Names
Anti-Cas9, Anti-CRISPR, Anti-CRISPR/Cas9, CRISPR antibody, Cas9 antibody

Application
WB: (1:2000 - 1:5000), ELISA: (1:5000-1:10000), ChIP or IP: (2 µg/106 cells), IF: (1:200-1:500), ICC: (1:200-1:500) 

User Guide & MSDS

[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

Product Citations

Kleinjan DA et. al. (October 2017). Drug-tunable multidimensional synthetic gene control using inducible degron-tagged dCas9 effectors. Nat Commun. 8(1):1191.

Yao L et. al. (December 2015). Multiple Gene Repression in Cyanobacteria Using CRISPRi. ACS Synth Biol.

Customer Reviews

Rating by r***@stanford.edu Verified Purchase Reviewed on: Monday 27 April, 2015
Application Description
I’ve tested the polyclonal Cas9 antibody. Results are attached

Procedural Details
K562 cells were nucleofected with pX330-U6-Chimeric_BB-CBh-hSpCas9 (Addgene plasmid #42230) and protein was harvested 24 hrs after nucleofections. Protein samples equivalent of approx. 25,000 cells was added 4x Laemmli Sample Buffer containing 1/10 DTT. Samples were boiled for 5 min, centrifuged at 16,300xg for 30s, and loaded onto an ‘Any kD Mini-PROTEAN TGX’ precast polyacrylamide gel (Bio-Rad), which was run at 43 mA for approx. 30 min. Protein was transferred to a PVDF membrane using the Trans-Blot Turbo Transfer System (Bio-rad) using the Bio-Rad program ‘HIGH MW’. After blotting, the membrane was washed once in TBS-T and blocked for 1 hr in TBS-T with 5% milk. The membrane was then washed three times in TBS-T and incubated overnight at 4 °C with primary antibody (cat. # A-1111) diluted 1:5,000 in TBS-T w. 0.5% milk. The membrane was then washed three times in TBS-T and incubated with secondary antibody (anti-rabbit HRP, Santa Cruz Biotechnology cat. # sc-2004) at 1:10,000 in TBS-T with 0.5% milk for 1 hr at room temperature. The blot was then washed five times in TBS-T and incubated with 2mL of ECL substrate (Bio-Rad, Clarity ECL Western Blotting Substrate) at room temperature for 5 min. The blot was visualized using the ChemiDoc XRS+ System (Bio-Rad)


Cas9 pAb western blot
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