CRISPR/Cas9 Antibodies
Cas9 is an RNA-guided DNA endonuclease from the microbial adaptive immune system CRISPR (clustered regularly interspaced short palindromic repeats) involved in gene editing and gene therapy. We offer antibodies to detect CRISPR-Cas9 that have been cited in 15+ publications with 5+ customer reviews. Browse our extensive catalog for CRISPR-Cas9 polyclonal antibodies, monoclonal antibodies, and a recombinant rabbit monoclonal antibody. Validated research applications for CRISPR/Cas9 antibodies include Chromatin Immunoprecipitation (ChIP), ELISA, ICC, Immunofluorescence, IHC, IHC-Paraffin, Flow Cytometry, IP, Simple Western, and Western Blot. Choose the anti-CRISPR-Cas9 antibody best suited for your research needs by viewing the technical details, images, references, reviews and related products. Each CRISPR-Cas9 antibody is fully covered by our Guarantee+.
Alternate Names for CRISPR/Cas9 Antibodies
anti-CRISPR/Cas9 antibody, anti-CRISPR antibody, anti-CRISPR-associated protein 9 nuclease antibody
17 Results for "crispr-cas9" in Primary Antibodies
![Simple Western: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - Image shows a specific band for Cas9 (observed molecular weight ~158 kDa) in HeLa Cas9 lysate but not in Hela WT lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-7A9-3A3-N-Terminus-Simple-Western-NBP2-36440-img0013.jpg "Simple Western: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - Image shows a specific band for Cas9 (observed molecular weight ~158 kDa) in HeLa Cas9 lysate but not in Hela WT lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.")
![Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - Hela cells were transiently transfected with an N-terminally Flag-tagged S. pyogenes Cas9 expression vector. The cells were stained with the Cas9 antibody followed by anti mouse-AF488 coupled secondary antibody. Nuclei were counter-stained with Hoechst 33342.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-7A9-3A3-N-Terminus-Immunocytochemistry-Immunofluorescence-NBP2-36440-img0002.jpg)
![Immunoprecipitation: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - HEK293T expressing N-terminally Flag-tagged S.pyogenes Cas9 were lysed 72h post transfection by resuspending the cells in Hunt buffer and subjecting to 3 freeze-thaw cycles in liquid nitrogen/ice. Proteins were immunoprecipitated from 100ug of whole cell lysate for 1h at 4C with Cas9 supernatant followed by incubation for 1h at 4C with a 1:1 mixture of protein A/G sepharose beads, or for 2h at 4C with Cas9 ab crosslinked to a 1:1 mixture of protein A/G sepharose beads. Beads were washed 2x with Hunt buffer and 1x with TBS. Bound proteins were eluted by boiling in Laemmli, separated by SDS-PAGE and transferred to nitrocellulose. Membrane was blocked, incubated with Cas9 ab, incubated with HRP anti-mouse secondary. *IgG heavy chain](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-7A9-3A3-N-Terminus-Immunoprecipitation-NBP2-36440-img0011.jpg)
![Western Blot: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - Analysis of lysate from Cas9 transfected HEK-293T cells using Cas9 antibody clone 7A9-3A3 at 2ug/ml concentration. The signal was developed using HRP-labelled anti-mouse secondary antibody and ECL based detection. Observed molecular weight is ~158 kDa.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-7A9-3A3-N-Terminus-Western-Blot-NBP2-36440-img0008.jpg)
![Western Blot: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - Human HEK293 cells were transfected with the indicated CRISPR-Cas9 plasmids, T2 guide RNA, and GFP transgene donor with homology arms to the AAVS1 targeted locus. HDR-mediated transgene integration was measured by FACS analysis of GFP-positive cells, resulting from targeted GFP transgene integration. Indels at the cleavage site were measured by the T7E1 assay. Asterisks indicate that the difference is statistically significant when comparing Cas9-HE, Cas9-HE-Geminin, and Cas9-Geminin to CRISPR-Cas9 in t-test (*P<0.05 or **P<0.005). Relative expression levels of and other fusions were analyzed by western blot with CRISPR-Cas9 and control anti-tubulin antibodies. Protein extracts were obtained with lysis buffer containing 150 mM NaCl. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-018-03475-7) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-7A9-3A3-N-Terminus-Western-Blot-NBP2-36440-img0016.jpg)
![Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - DF-1 stable cell line (chicken fibroblast) expressing Cas9 (left )or wildtype DF-1 cells (right) stained with Cas9 antibody NBP2-36440 and phalloidin and DAPI to visualise F-actin and DNA respectively. Cells fixed with 4% PFA. Antibody at 1:500 overnight at 4C. ICC/IF image submitted by a verified customer review.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-7A9-3A3-N-Terminus-Immunocytochemistry-Immunofluorescence-NBP2-36440-img0014.jpg)
![Immunohistochemistry-Frozen: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - Analysis of a formalin fixed 20um thick frozen section of mouse brain with GBM xenograft tumor areas (GBM cells over expressing SpyCas9 through lentivirus infection). CRISPR-Cas9 antibody (clone 7A9-3A3) was used at 1:50 dilution. The signal was detected using immunofluoresence labeled secondary antibody via Confocal microscopy. Image submitted via verified customer review.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-7A9-3A3-N-Terminus-Immunohistochemistry-Frozen-NBP2-36440-img0010.jpg)
![Western Blot: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - 20 ug whole cell lysates from control MEF (MEF-WT) and MEF-Cas9 stable cell line. CRISPR-Cas9 antibody (clone 7A9-3A3) was used at 1:1000 dilution. Observed molecular weight is ~158 kDa. Image submitted via verified customer review.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-7A9-3A3-N-Terminus-Western-Blot-NBP2-36440-img0012.jpg)
![Western Blot: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - Chicken fibroblasts DF-1 cells stably expressing Cas9 in lanes 2 and 4, wild type fibroblasts in lanes 1 and 3 blot, probed with Cas9 antibody. WB image submitted by a verified customer review.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-7A9-3A3-N-Terminus-Western-Blot-NBP2-36440-img0015.jpg)
![Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - Analysis of Crispr-Cas9 transfected HEK293 cells using CRISPR-Cas9 antibody (clone 7A9-3A3). Red staining represents CRISPR-Cas9 positivity while DAPI stained nuclei are visible in blue color. Image submitted via verified customer review.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-7A9-3A3-N-Terminus-Immunocytochemistry-Immunofluorescence-NBP2-36440-img0007.jpg)
![Immunoprecipitation: Mouse Monoclonal CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus [NBP2-36440] - LC-MS/MS signals of a surrogate peptide from Cas9 and a Cas9 variant after immunoprecipitation using CRISPR-Cas9 Antibody (7A9-3A3) [Biotin] (NBP2-36440B). Image from a verified customer review.](http://images.novusbio.com/fullsize/antibody/nbp2-36440_mouse-monoclonal-crispr-cas9-antibody-7a9-3a3-n-terminus-immunoprecipitation-1582024135956..png)
![Western Blot: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - BSA Free [NBP2-36440] - Identification of “HDR-enhancer” (HE) domain of CtIP. a Schematic diagram of CtIP protein showing different truncated CtIP proteins have been fused to Cas9 & tested for their ability to stimulate HDR. Various sequence features of CtIP, including tetramerization & dimerization domains, & CDK phosphorylation sites S233, T245, & S276, are indicated. b Identification of a domain of CtIP, called HE, spanning aa 1 to 296, which is able to stimulate HDR when fused to Cas9. Human RG37 fibroblasts transfected w/ indicated plasmids expressing Cas9 or Cas9–CtIP derivatives, T2 guide RNA plasmid, & GFP transgene donor w/ homology arms to targeted AAVS1 locus. Expression of fusion proteins examined by WB (Supplementary Fig. 1). Data are from four independent experiments. Error bars indicate standard deviation. c Functional analysis of HE domain. HEK293 cells transfected w/ indicated Cas9 plasmids, T2 guide RNA, & GFP transgene donor w/ homology arms to AAVS1 targeted locus. HDR-mediated transgene integration measured by FACS analysis of GFP-positive cells, resulting from targeted GFP transgene integration. Indels at cleavage site measured by T7E1 assay. Results are expressed as mean of relative HDR or indel frequencies calculated by normalizing every HDR or indel frequency by induced by Cas9, respectively. Asterisks indicate difference is statistically significant when comparing Cas9–CtIP or Cas9–HE derivatives to Cas9 in nonparametric t-test (*P<0.05, **P<0.005, or ***P<0.0005). Data are from four independent experiments. Error bars indicate standard deviation. The relative expression levels of Cas9 & Cas9–HE derivatives analyzed by WB using anti-Cas9 & control anti-tubulin antibodies Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/29556040), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp2-36440_mouse-monoclonal-crispr-cas9-antibody-7a9-3a3-n-terminus-310202416114788.jpg)
![Western Blot: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - BSA Free [NBP2-36440] - Stimulation of transgene integration by Cas9–HE & Cas9–Geminin. Relative frequencies of HDR & indels induced by Cas9 or fusion of Cas9 to HE domain, Geminin degron, or both. Human HEK293 cells were transfected with the indicated Cas9 plasmids, T2 guide RNA, & GFP transgene donor with homology arms to the AAVS1 targeted locus. HDR-mediated transgene integration was measured by FACS analysis of GFP-positive cells, resulting from targeted GFP transgene integration. Indels at the cleavage site were measured by the T7E1 assay. The results are expressed as the mean of relative HDR or indel frequency calculated by normalizing every HDR or indel frequency by that induced by Cas9. Asterisks indicate that the difference is statistically significant when comparing Cas9–HE, Cas9–HE–Geminin, & Cas9–Geminin to Cas9 in t-test (*P<0.05 or **P<0.005). Data are from three independent experiments. Error bars indicate standard deviation. Relative expression levels of Cas9 & other fusions were analyzed by western blot with anti-Cas9 & control anti-tubulin antibodies. Protein extracts were obtained with lysis buffer containing 150 mM NaCl, which resulted in inefficient solubilization of Cas9 fusions with the HE domain compared to those of Cas9 & Cas9–Geminin Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29556040), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp2-36440_mouse-monoclonal-crispr-cas9-antibody-7a9-3a3-n-terminus-3102024161361.jpg)
![Western Blot: CRISPR-Cas9 Antibody (6G12) - C-terminus [NBP2-52398] - CRISPR-Cas9 Antibody (6G12) - C-Terminus [NBP2-52398] - Control HeLa cells (un-transfected) and HeLa cells expressing Flag-tagged S. pyogenes's CRISPR-Cas9 under the control of PTight (Tet-ON) promoter. Samples were treated for 24 hours with 1ug/uL of Doxycyclin and lysed under native conditions. 30 ug of the whole cell lysate from each sample type per lane was separated by 7.5% SDS-PAGE. Nitrocellulose membrane was incubated with CRISPR-Cas9 antibody clone 6G12 (hybridoma supernatant diluted 1:100 at 4C O/N). After washing, the membranes were incubated with secondary HRP-coupled antibody and bands were visualized by ECL and exposure of X-ray films. Prestained marker bands were visualized with Blue Marker Antibody (NBP2-33376). The image shown is from 1 minute exposure time. Observed molecular weight is ~158 kDa.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-Terminus-Western-Blot-NBP2-52398-img0008.jpg "Western Blot: CRISPR-Cas9 Antibody (6G12) - C-terminus [NBP2-52398] - CRISPR-Cas9 Antibody (6G12) - C-Terminus [NBP2-52398] - Control HeLa cells (un-transfected) and HeLa cells expressing Flag-tagged S. pyogenes's CRISPR-Cas9 under the control of PTight (Tet-ON) promoter. Samples were treated for 24 hours with 1ug/uL of Doxycyclin and lysed under native conditions. 30 ug of the whole cell lysate from each sample type per lane was separated by 7.5% SDS-PAGE. Nitrocellulose membrane was incubated with CRISPR-Cas9 antibody clone 6G12 (hybridoma supernatant diluted 1:100 at 4C O/N). After washing, the membranes were incubated with secondary HRP-coupled antibody and bands were visualized by ECL and exposure of X-ray films. Prestained marker bands were visualized with Blue Marker Antibody (NBP2-33376). The image shown is from 1 minute exposure time. Observed molecular weight is ~158 kDa.")
![<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody (6G12) - C-terminus [NBP2-52398] - CRISPR-Cas9 Antibody (6G12) - C-Terminus [NBP2-52398] - HeLa cells or HeLa cells expressing Flag-tagged SpCas9 under the control of the PTight (Tet-ON) promoter were treated for 24h with 1ug/uL Doxycyclin, fixed and permeabilized with Methanol/Acetone and blocked in 2% BSA in PBS for 2 hours at RT. Cells were stained with 6G12 hybridoma supernatant at 1:10 at 4C O/N, followed by incubation with anti mouse-Alexa Fluor 488 coupled secondary antibody for 1h at RT. Nuclei were counter-stained with Hoechst 33342.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-Terminus-Immunocytochemistry-Immunofluorescence-NBP2-52398-img0011.jpg)
![Simple Western: CRISPR-Cas9 Antibody (6G12) - C-terminus [NBP2-52398] - Image shows a specific band for Cas9 in 0.2 mg/mL of HeLa Cas9 lysate but not in Hela WT lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Observed molecular weight is ~158 kDa.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-terminus-Simple-Western-NBP2-52398-img0014.jpg)
![<b>Independent Antibodies Validation. </b>Chromatin Immunoprecipitation: CRISPR-Cas9 Antibody (6G12) - C-terminus [NBP2-52398] - CRISPR-Cas9 Antibody (6G12) - C-Terminus [NBP2-52398] - CRISPR-Cas9 Antibody (6G12) [NBP2-52398] - NIH3T3 cells stably expressing GFP-H2B, nuclease dead Cas9, and a GFP-targeting gRNA were fixed with formaldehyde, harvested and sonicated to get 200-500bp DNA fragments. 50ug chromatin was incubated over night at 4C with the indicated antibodies (200ul hybridoma SN, 5ug anti-Flag [M2, Sigma]) followed by incubation with protein G beads for 3h at 4C. After washing chromatin was eluted from the beads and crosslinking was reversed over night at 65C. After a proteinase K digestion step, DNA was separated using phenol/chloroform/isoamyl alcohol, precipitated with ethanol/sodium acetate and dissolved in water. For qPCR, primers either targeting the GFP gene or as negative control non-targeted regions (Ppap2c +7122 and Prkcd +24069 from transcription start) were used.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-Terminus-Chromatin-Immunoprecipitation-ChIP-NBP2-52398-img0013.jpg)
![Western Blot: CRISPR-Cas9 Antibody (6G12) - C-terminus [NBP2-52398] - CRISPR-Cas9 Antibody (6G12) - C-Terminus [NBP2-52398] - Whole cell protein from 293T cells transfected with Cas9-Flag (~150 kDa) was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2 ug/mL anti-Cas9 (6G12) in 1% milk, and detected with an anti-mouse HRP secondary antibody using chemiluminescence.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-Terminus-Western-Blot-NBP2-52398-img0009.jpg)
![Immunoprecipitation: CRISPR-Cas9 Antibody (6G12) - C-terminus [NBP2-52398] - CRISPR-Cas9 Antibody (6G12) - C-Terminus [NBP2-52398] - HEK293 cells expressing Flag-SpCas9 were lysed under native conditions. SpCas9 was immunoprecipitated at 4C from 300 ug of whole cell lysate with the 6G12 antibody and a 1:1 mixture of protein A/G sepharose. After 4x washing, the bound proteins were boiled off the beads, separated by 7.5% SDS-PAGE and transfered to nitrocellulose membranes, and SpCas9 was detected with a rabbit polyclonal Cas9 antibody. After washing, the membranes were incubated with secondary HRP-coupled antibody and bands were visualized by ECL and exposure of X-ray films.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-Terminus-Immunoprecipitation-NBP2-52398-img0012.jpg)
![Western Blot: CRISPR-Cas9 Antibody (6G12) - C-terminus - Azide and BSA Free [NBP2-80680] - Whole cell protein from 293T cells transfected with Cas9-Flag (~150 kDa) was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2 ug/mL anti-Cas9 (6G12) in 1% milk, and detected with an anti-mouse HRP secondary antibody using chemiluminescence. Image from the standard format of this antibody.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-terminus-Azide-and-BSA-Free-Western-Blot-NBP2-80680-img0002.jpg "Western Blot: CRISPR-Cas9 Antibody (6G12) - C-terminus - Azide and BSA Free [NBP2-80680] - Whole cell protein from 293T cells transfected with Cas9-Flag (~150 kDa) was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2 ug/mL anti-Cas9 (6G12) in 1% milk, and detected with an anti-mouse HRP secondary antibody using chemiluminescence. Image from the standard format of this antibody.")
![Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody (6G12) - C-terminus - Azide and BSA Free [NBP2-80680] - HeLa cells or HeLa cells expressing Flag-tagged SpCas9 under the control of the PTight (Tet-ON) promoter were treated for 24h with 1ug/uL Doxycyclin, fixed and permeabilized with Methanol/Acetone and blocked in 2% BSA in PBS for 2 hours at RT. Cells were stained with 6G12 hybridoma supernatant at 1:10 at 4C O/N, followed by incubation with anti mouse-Alexa Fluor 488 coupled secondary antibody for 1h at RT. Nuclei were counter-stained with Hoechst 33342. Image from the standard format of this antibody.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-terminus-Azide-and-BSA-Free-Immunocytochemistry-Immunofluorescence-NBP2-80680-img0003.jpg)
![Chromatin Immunoprecipitation: CRISPR-Cas9 Antibody (6G12) - C-terminus - Azide and BSA Free [NBP2-80680] - NIH3T3 cells stably expressing GFP-H2B, nuclease dead Cas9, and a GFP-targeting gRNA were fixed with formaldehyde, harvested and sonicated to get 200-500bp DNA fragments. 50ug chromatin was incubated over night at 4C with the indicated antibodies (200ul hybridoma SN, 5ug anti-Flag [M2, Sigma]) followed by incubation with protein G beads for 3h at 4C. After washing chromatin was eluted from the beads and crosslinking was reversed over night at 65C. After a proteinase K digestion step, DNA was separated using phenol/chloroform/isoamyl alcohol, precipitated with ethanol/sodium acetate and dissolved in water. For qPCR, primers either targeting the GFP gene or as negative control non-targeted regions (Ppap2c +7122 and Prkcd +24069 from transcription start) were used. Image from the standard format of this antibody.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-terminus-Azide-and-BSA-Free-Chromatin-Immunoprecipitation-NBP2-80680-img0004.jpg)
![Western Blot: CRISPR-Cas9 Antibody (6G12) - C-terminus - Azide and BSA Free [NBP2-80680] - Control HeLa cells (un-transfected) and HeLa cells expressing Flag-tagged S. pyogenes's CRISPR-Cas9 under the control of PTight (Tet-ON) promoter. Samples were treated for 24 hours with 1ug/uL of Doxycyclin and lysed under native conditions. 30 ug of the whole cell lysate from each sample type per lane was separated by 7.5% SDS-PAGE. Nitrocellulose membrane was incubated with CRISPR-Cas9 antibody clone 6G12 (hybridoma supernatant diluted 1:100 at 4C O/N). After washing, the membranes were incubated with secondary HRP-coupled antibody and bands were visualized by ECL and exposure of X-ray films. Prestained marker bands were visualized with Blue Marker Antibody (NBP2-33376). The image shown is from 1 minute exposure time. Observed molecular weight is ~158 kDa. Image from the standard format of this antibody.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-terminus-Azide-and-BSA-Free-Western-Blot-NBP2-80680-img0001.jpg)
![Simple Western: CRISPR-Cas9 Antibody (6G12) - C-terminus - Azide and BSA Free [NBP2-80680] - Image shows a specific band for Cas9 in 0.2 mg/mL of HeLa Cas9 lysate but not in Hela WT lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system. Observed molecular weight is ~158 kDa. Image from the standard format of this antibody.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-terminus-Azide-and-BSA-Free-Simple-Western-NBP2-80680-img0006.jpg)
![Immunoprecipitation: CRISPR-Cas9 Antibody (6G12) - C-terminus - Azide and BSA Free [NBP2-80680] - HEK293 cells expressing Flag-SpCas9 were lysed under native conditions. SpCas9 was immunoprecipitated at 4C from 300 ug of whole cell lysate with the 6G12 antibody and a 1:1 mixture of protein A/G sepharose. After 4x washing, the bound proteins were boiled off the beads, separated by 7.5% SDS-PAGE and transfered to nitrocellulose membranes, and SpCas9 was detected with a rabbit polyclonal Cas9 antibody. After washing, the membranes were incubated with secondary HRP-coupled antibody and bands were visualized by ECL and exposure of X-ray films. Image from the standard format of this antibody.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6G12-C-terminus-Azide-and-BSA-Free-Immunoprecipitation-NBP2-80680-img0005.jpg)
![Western Blot: CRISPR-Cas9 Antibody [NBP2-52717] - HEK293T were transfected with either WT CAS9 plasmid, CAS9 nickase plasmid or control vector. Equivalent amount of each cell lysates (10ug per lane) were immunoblotted with (1:2000). Theoretical molecular weight ~158 kDa.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-Western-Blot-NBP2-52717-img0002.jpg "Western Blot: CRISPR-Cas9 Antibody [NBP2-52717] - HEK293T were transfected with either WT CAS9 plasmid, CAS9 nickase plasmid or control vector. Equivalent amount of each cell lysates (10ug per lane) were immunoblotted with (1:2000). Theoretical molecular weight ~158 kDa.")
![Western Blot: CRISPR-Cas9 Antibody [NBP2-52717] - A. After the induction of CAS9 and XCAS9 in 293 cells with 2 ug/mL doxycycline for 3 days, CAS9 expression was accessed by WB. XCAS9 is a CAS9 variant with potentially higher fidelity and broader compatibility. B) The impact of expression levels of CAS9 on DNA DSB damage in hESCs. At the same lentiviral titers, the expression levels of CAS9 in hESCs transduced by standard lentiviral vector are higher than those transduced by the inducible lentiviral vector after 2 ug/mL Doxy treatment. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32170574) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-Western-Blot-NBP2-52717-img0007.jpg)
![Immunoprecipitation: CRISPR-Cas9 Antibody [NBP2-52717] - HEK293T were transfected with either WT CAS9 plasmid, CAS9 nickase plasmid (GE100019), or negative control vector (PS100001). Input: 10ug total cell lysates; WB was performed (1:2000). IP: 100ug of cell lysates were precipitated with 20ug of NBP2-52717 or control antibody, and 5ug Protein G Beads. Mouse anti-DDK monoclonal antibody was used in WB to detect the immuno-precipitated Cas9 (as Cas9 is Myc-DDK tagged)](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-Immunoprecipitation-NBP2-52717-img0006.jpg)
![Western Blot: CRISPR-Cas9 Antibody [NBP2-52717] - The expression of CAS9 in hiPSCs & hESCs promotes DNA DSB damage. (A) The inducible expression of CAS9 promotes DNA DSB damage responses in hiPSCs after 2 µg/mL Doxy treatment. The relative levels of the phosphorylation of p53 & H2AX are indicated at the bottom. Consistent data were obtained from two independent experiments. (B) The impact of expression levels of CAS9 on DNA DSB damage in hESCs. At the same lentiviral titers, the expression levels of CAS9 in hESCs transduced by standard lentiviral vector are higher than those transduced by the inducible lentiviral vector after 2 µg/mL Doxy treatment. Much lower expression levels of CAS9 can also promote DNA DSB damage in hESCs after the treatment with lower dosages of Doxy. The relative levels of the phosphorylation of p53 & H2AX are indicated at the bottom Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32170574), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp2-52717_rabbit-crispr-cas9-pab-31020241612195.jpg)
![Western Blot: CRISPR-Cas9 Antibody [NBP2-52717] - dCAS9 & Cpf1 impair NHEJ & induce genetic mutations. (A) Co-immunoprecipitation assay confirmed the interaction between dCAS9 & KU86. (B) Comet assay analysis of DNA damage in hESCs expressing dCAS9 or treated with doxorubicin. CTL, human fibroblasts with lentiviral empty vector were treated with 2 µg/mL doxycycline for three days; Doxy, Dox, Doxy + Dox, human fibroblasts with lentiviral CAS9 inducible expression vector were treated with 2 µg/mL doxycycline for 3 days or 0.5 µmol/L Dox for 2 h or 2 µg/mL doxycycline for three days + 0.5 µmol/L Dox for 2 h, respectively. Tail length was analyzed using Image J software. Data are represented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. (C) The expression of dCAS9 induces mutation of endogenous HPRT gene. WT, WT hESCs; CTL, CAS9, dCAS9, hESCs with empty expression vector, CAS9 inducible expression vector. Cells with dCAS9 inducible expression vector were treated with 2 µg/mL doxycycline for 3 days before HAT treatment. n = 3. Data are presented as mean value ± SD. **P < 0.01, ***P < 0.001. (D) The expression of Cpf1 increased the levels of gamma H2AX. (E) Cpf1 interacts with KU86 as confirmed by Co-immunoprecipitation. Protein extract of Flag-tagged Cpf1 was immunoprecipitated with anti-Flag antibody & the presence of Cpf1 & KU86 in the immunoprecipitate was examined by Western blot Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32170574), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp2-52717_rabbit-crispr-cas9-pab-310202416113547.jpg)
![Western Blot: CRISPR-Cas9 Antibody [NBP2-52717] - Both CAS9 & XCAS9 impair NHEJ & induce genetic mutations. (A & B) Expression of CAS9 & XCAS9 impairs NHEJ. Traffic Light Reporter system was established in 293 cells harboring the CAS9 & XCAS9 inducible expression vectors. After the induction of CAS9 & XCAS9 expression with 2 µg/mL doxycycline for 3 days (left panel), the efficiency of NHEJ (mcherry) & HDR (GFP) was analyzed by flow cytometry (middle panel). Statistic analysis of the efficiency of NHEJ (left panel of A) & HDR (B) is presented. n = 3. Data are presented as mean values ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. ns, non-significant. (C) The expression of CAS9 in hESCs induces genomic mutations at the endogenous HPRT locus. After hESCs harboring CAS9 inducible expression vector were selected with HAT medium for 5 days, they were treated with 2 µg/mL doxycycline for CAS9 expression for various time periods, & subsequently, treated with 5 µg/mL 6-TG or mock treated for 4 days. Mutational rate is calculated as the ratio of colony number in 6-TG treated samples versus untreated controls. n = 3. Data are presented as mean values ± SD. ***P < 0.001. (D) XCAS9 interacts with KU86. Protein extracts from 293FT cells expressing Flag-tagged CAS9 or XCAS9 were immunoprecipitated with anti-Flag antibody. The immune precipitates were analyzed for the presence of CAS9, XCAS9 & KU86. The relative ratio of KU86 versus CAS9 or XCAS9 is indicated. (E) The expression of XCAS9 increases the number of gamma H2AX foci in hESCs. hESCs harboring XCAS9 inducible expression vector were treated with or without 2 µg/mL doxycycline for 3 days. n = 20. Scale bar, 10 µm. Data are presented as mean values ± SD. ***P < 0.01 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32170574), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp2-52717_rabbit-crispr-cas9-pab-310202416114779.jpg)
![Western Blot: CRISPR-Cas9 Antibody [NBP2-52717] - The expression of CAS9 in human fibroblasts promotes DNA DSB damage & activates DNA damage response pathways. (A) The expression of CAS9 in human fibroblasts activates DNA damage responses. The expression of CAS9 was induced with 2 µg/mL Doxy treatment. The relative levels of phosphorylation of BRCA1, CHK1, CHK2 & p53 are indicated at the bottom. n = 3. Data are presented as mean value ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. (B) The expression of CAS9 increased the number of gamma H2AX foci in human fibroblasts. CTL or CAS9, human fibroblasts with CAS9 inducible expression cassette plated on chamber slides were treated with or without 2 µg/mL doxycycline for 3 days. The expression of CAS9 & gamma H2AX foci was revealed by immunoflourescence analysis. Representative images are shown. Scale bar, 10 µm. Unpaired t test. n = 20. Data are presented as mean values ± SD. ***P < 0.001. (C) CAS9 induces DNA DSB damage in human fibroblasts. CTL, human fibroblasts with lentiviral empty vector were treated with 2 µg/mL doxycycline for three days; Doxy, Dox, Doxy + Dox, human fibroblasts with lentiviral CAS9 inducible expression vector were treated with 2 µg/mL doxycycline for 3 days or 0.5 µmol/L Dox for 2 h or 2 µg/mL doxycycline for three days + 0.5 µmol/L Dox for 2 h, respectively. Representative images are shown. n = 40. Unpaired t test. Data are presented as mean value ± SD. **P < 0.01 Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32170574), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp2-52717_rabbit-crispr-cas9-pab-310202416121950.jpg)
![Western Blot: CRISPR-Cas9 Antibody (6H4) [NBP2-81124] - Total protein from mock and Cas9 transfected 293 cells, HeLa and 3T3 cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 0.5 ug/ml anti-Cas9 in block buffer and detected with an anti-mouse HRP secondary antibody using West Pico PLUS chemiluminescence detection reagent. Theoretical molecular weight: 158.4 kDa.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-6H4-Western-Blot-NBP2-81124-img0001.jpg "Western Blot: CRISPR-Cas9 Antibody (6H4) [NBP2-81124] - Total protein from mock and Cas9 transfected 293 cells, HeLa and 3T3 cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 0.5 ug/ml anti-Cas9 in block buffer and detected with an anti-mouse HRP secondary antibody using West Pico PLUS chemiluminescence detection reagent. Theoretical molecular weight: 158.4 kDa.")
![Western Blot: CRISPR-Cas9 Antibody [NBP3-05547] - Western blot analysis of HEK293 cell lysates using chicken pAb to CRISPR-Cas9 [1] protein standard (red), [2] non-transfected cells and [3] transfected cells with GFP-CRISPR-Cas9 (C-terminal 803-1053 amino acids of CRISPR-Cas9) fusion construct. The band at about 53kDa corresponds to the GFP-CAS9 fusion protein.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-Western-Blot-NBP3-05547-img0002.jpg "Western Blot: CRISPR-Cas9 Antibody [NBP3-05547] - Western blot analysis of HEK293 cell lysates using chicken pAb to CRISPR-Cas9 [1] protein standard (red), [2] non-transfected cells and [3] transfected cells with GFP-CRISPR-Cas9 (C-terminal 803-1053 amino acids of CRISPR-Cas9) fusion construct. The band at about 53kDa corresponds to the GFP-CAS9 fusion protein.")
![Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody [NBP3-05547] - HEK293 cells were transfected with a construct including the N-terminal 608 amino acids of S. aureus CRISPR-Cas9 fused to GFP and stained with CRISPR-Cas9 Antibody in red. Transfected cells express the green fusion protein and bind the antibody in red, producing a yellow signal. Nuclear DNA in transfected and non-transfected cells is revealed with the blue DNA stain DAPI.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-Immunocytochemistry-Immunofluorescence-NBP3-05547-img0001.jpg)
![Western Blot: CRISPR-Cas9 Antibody [NBP2-66379] - Total protein from Cas9 transfected Hek293T cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-Cas9 in block buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Expected molecular weight: 158.4 kDa.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-Western-Blot-NBP2-66379-img0001.jpg "Western Blot: CRISPR-Cas9 Antibody [NBP2-66379] - Total protein from Cas9 transfected Hek293T cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-Cas9 in block buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. Expected molecular weight: 158.4 kDa.")
![Western Blot: CRISPR-Cas9 Antibody [NBP3-05548] - Western blot analysis of HEK293 cell lysates using rabbit pAb to S. pyogenes CRISPR-Cas9 Antibody: [1] protein standard (red), [2] non-transfected cells, [3] transfected cells with C-terminal (814-1372 amino acids) of S. pyogenes CRISPR-Cas9, and [4] transfected HEK293 cells with N-terminal (1-608 amino acids) of S. pyogenes CRISPR-Cas9. 60kDa and 68kDa bands correspond to S. pyogenes CRISPR-Cas9 C-terminal and N-terminal constructs respectively.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-Western-Blot-NBP3-05548-img0002.jpg "Western Blot: CRISPR-Cas9 Antibody [NBP3-05548] - Western blot analysis of HEK293 cell lysates using rabbit pAb to S. pyogenes CRISPR-Cas9 Antibody: [1] protein standard (red), [2] non-transfected cells, [3] transfected cells with C-terminal (814-1372 amino acids) of S. pyogenes CRISPR-Cas9, and [4] transfected HEK293 cells with N-terminal (1-608 amino acids) of S. pyogenes CRISPR-Cas9. 60kDa and 68kDa bands correspond to S. pyogenes CRISPR-Cas9 C-terminal and N-terminal constructs respectively.")
![Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody [NBP3-05548] - HEK293 cells were transfected with a construct including the N-terminal 608 amino acids of S. pyogenes CRISPR-Cas9 fused to GFP and stained with CRISPR-Cas9 Antibody in red. Transfected cells express the green fusion protein and bind the antibody in red, producing a yellow signal. Nuclear DNA in transfected and non-transfected cells is revealed with the blue DNA stain DAPI.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-Immunocytochemistry-Immunofluorescence-NBP3-05548-img0001.jpg)
![Western Blot: CRISPR-Cas9 Antibody (SR1104) [NBP3-21732] - Western blot analysis using NBP3-21732 in (1) 293T cell lysate transfected with CRISPR-Cas9; (2) 293T cell lysate; (3) 3T3 cell lysate; (4) PC12 cell lysate.](http://images.novusbio.com/fullsize/nbp3-21732_rabbit-crispr-cas9-mab-sr1104-6720231051265.jpg "Western Blot: CRISPR-Cas9 Antibody (SR1104) [NBP3-21732] - Western blot analysis using NBP3-21732 in (1) 293T cell lysate transfected with CRISPR-Cas9; (2) 293T cell lysate; (3) 3T3 cell lysate; (4) PC12 cell lysate.")
![Western Blot: CRISPR-Cas9 Antibody (JM11-55) [NBP2-67000] - Analysis of CRISPR-Cas9 SP on 293T transfected with Cas9 cell lysates with Rabbit anti-CRISPR-Cas9 SP antibody at 1/1,000 dilution.Lysates/proteins at 20 ug/Lane.Predicted band size: 158 kDaObserved band size: 158 kDaExposure time: 25 seconds; ECL: K1801;4-20% SDS-PAGE gel.Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.](http://images.novusbio.com/fullsize/nbp2-67000_rabbit-monoclonal-crispr-cas9-antibody-jm11-55-western-blot-317202491743..jpg "Western Blot: CRISPR-Cas9 Antibody (JM11-55) [NBP2-67000] - Analysis of CRISPR-Cas9 SP on 293T transfected with Cas9 cell lysates with Rabbit anti-CRISPR-Cas9 SP antibody at 1/1,000 dilution.Lysates/proteins at 20 ug/Lane.Predicted band size: 158 kDaObserved band size: 158 kDaExposure time: 25 seconds; ECL: K1801;4-20% SDS-PAGE gel.Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.")
![Western Blot: CRISPR-Cas9 Antibody [NBP2-52718] - Lane 1: 293T non transfected cell lysate. Lane 2: 293T Cas9 over expressed cell lysate. Load: 15ug per lane. Primary Antibody cas9 used at 1ug/mL overnight at 4C. Secondary Antibody: goat anti-rabbit HRP at 1:40,000 for 30 min at room temp. Expected theoretical molecular weight: 158 kDa.](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-Western-Blot-NBP2-52718-img0001.jpg "Western Blot: CRISPR-Cas9 Antibody [NBP2-52718] - Lane 1: 293T non transfected cell lysate. Lane 2: 293T Cas9 over expressed cell lysate. Load: 15ug per lane. Primary Antibody cas9 used at 1ug/mL overnight at 4C. Secondary Antibody: goat anti-rabbit HRP at 1:40,000 for 30 min at room temp. Expected theoretical molecular weight: 158 kDa.")
![Immunofluorescence: CRISPR-Cas9 Antibody [NBP2-52718] - HeLa cells transfected with GFP+Cas9 or GFP+CD-3. Fixation: Formaldehyde fixed, permeabilized Triton X-100. Primary: Anti-Cas9 used at 1ug/mL. Secondary: Donkey anti-Rabbit at 1ug/mL, staining seen in red. Bottom images show GFP stain overlay](http://images.novusbio.com/fullsize/CRISPR-Cas9-Antibody-Immunofluorescence-NBP2-52718-img0002.jpg)