Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468)
![Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468)](https://www.abcam.com/ps/products/191/ab191468/Images/ab191468-244896-anti-crispr-cas9-antibody-7a9-3a3-western-blot.jpg "Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468)")
Key features and details
- Mouse monoclonal [7A9-3A3] to CRISPR-Cas9
- Suitable for: ICC/IF, WB
- Reacts with: Streptococcus pyogenes
- Isotype: IgG1
Overview
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Product name
Anti-CRISPR-Cas9 antibody [7A9-3A3]
See all CRISPR-Cas9 primary antibodies -
Description
Mouse monoclonal [7A9-3A3] to CRISPR-Cas9 -
Host species
Mouse -
Specificity
Ab191468 detects Cas9 and dCas9. This product has not been tested against SaCas9. Please contact Abcam Scientific Support for more information.
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Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Streptococcus pyogenes -
Immunogen
Recombinant fragment corresponding to Streptococcus pyogenes CRISPR-Cas9 (N terminal).
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Positive control
- WB: S2 cells transfected with CRISPR-Cas9 ICC-IF: NIH/3T3-Cas9 transfected cells.
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General notes
This antibody clone is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Some batches contain 6.97% L-Arginine as a stabilizing agent. For lot-specific buffer information, please contact our Scientific Support team. -
Concentration information loading... -
Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
7A9-3A3 -
Isotype
IgG1 -
Light chain type
kappa
Images
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Western blot - Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468)All lanes : Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468) at 5 µg/ml
Lane 1 : S2 non-transfected cell lysate
Lane 2 : S2 cells transfected with CRISPR-Cas9 plasmid
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 160 kDa
Observed band size: 160 kDa
Exposure time: 4 minutesWe recommend using 3% milk as the blocking agent in Western Blot.
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![Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468)](https://www.abcam.com/ps/products/191/ab191468/Images/ab191468-272642-anti-crispr-cas9-antibody-7a9-3a3-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468)ab191468 stained in NIH3T3 cells. Untreated and Cas9 transfected cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab191468 at 10µg/ml and ab6046 (Rabbit polyclonal to beta tubulin) at 1ug/ml overnight at +4°C. The secondary antibodies were ab150117 (colored green) used at 1 ug/ml and ab150087 (pseudo-colored red) used at 2ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.
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![Western blot - Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468)](https://www.abcam.com/ps/products/191/ab191468/Images/ab191468-385393-AntiCRISPRCas9-antibody-7A93A3.jpg)
Western blot - Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468) This image was courtesy of an anonymous AbReviewLane 1: HeLa inducible Cas9 cell line in the absence of Doxcycline (-DOX)
Lane 2: HeLa inducible Cas9 cell line in the treated for 24 hours in the presence of 1ug/ml of Doxcycline (+DOX).
Blot was probed with Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468) 1:2000 and Anti-Tubulin Antibody DM1A (Sigma) as loading control. Shows specific band at 170kDA in Dox treated cells and tubulin band at 50kDa. Lysis and running Cells lysed in 1.5X Lammeli buffer +0.15M DTT syringed 10X with 25g needle and then boiled 100 degrees C for 10 minutes. 10ug of sample run out on 4-12% Bis Tris Nupage gel in 1X MOPS buffer WB protocol Western blot was performed by wet transfer of gel onto nitrocellulose membrane 100V for 2 h 4 degrees. Blocked in 5% milk PBST for 1 h room temp (1X PBS + 0.1% tween 20) incubated 24h at 4 degrees in 5% PBST with primary antibodies 1:2000 Anti-CRISPR-Cas9 antibody [7A9-3A3] (ab191468) as well as 1:3000 Anti Tubulin Mouse monoclonal DM1A antibody ( Sigma) Washed 3X PBST IgG anti mouse HRP 1:5000 (Jackson immunoscience) Washed 3x PBST Washed
HeLa cell line contains Cas9 under the control of a Doxcycline/Tetracycline inducible promotor (Tet on) Specific band at 170 kDa appears upon treatment with Doxcycline 1ug/ml for 24hours.1X PBS Developed using ECL prime kit (GE healthcare).
