CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free

Western Blot: CRISPR-Cas9 Antibody (7A9-3A3)N-TerminusAzide and BSA Free [NBP2-80679]

Western Blot: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679] - 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. Image from the standard format of this antibody.

Immunocytochemistry/ Immunofluorescence: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679]

Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679] - 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. Image from the standard format of this antibody.

Simple Western: CRISPR-Cas9 Antibody (7A9-3A3)N-TerminusAzide and BSA Free [NBP2-80679]

Simple Western: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679] - 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. Image from the standard format of this antibody.

Immunoprecipitation: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679]

Immunoprecipitation: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679] - 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 Image from the standard format of this antibody.

Western Blot: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - Azide and BSA Free [NBP2-80679] -

Cas9-dependent breaks are invisible to DNA end resection enzymes. See also Supplementary Fig. 1.a Sequence of a DNA segment within the ~2.4 kbp circular plasmid-based DNA substrate used for the Cas experiments. The sequence of the crRNAs is shown in bold for both Cas9 and Cas12a. The PAM sequence is underlined, and the arrowheads indicate the putative cleavage sites. The sequence in green corresponds to the radioactive probe that anneals to the substrates if resection has taken place. The red asterisk indicates the position of the radioactive label. b Representative agarose gel electrophoresis of the substrates treated with the indicated Cas variants. The separation was performed on a 1% TAE gel in the presence of GelRed. The position of the various products is indicated. DA: Cas9 D10A; HA: Cas9 H840A; d: catalytically inactive Cas9; Fn: Francisella novicida Cas12a; Mb: Moraxella bovoculi Cas12a. A representative of two independent experiments is shown. c Top, a schematic overview of the assay. Bottom, representative annealing DNA end resection assay showing resection by the MRE11 complex (25 nM Mre11-Rad50 with 200 nM phosphorylated Sae2, MR-pSae2) of plasmid-based DNA substrate treated with the indicated Cas variants. The probe anneals 92 bp away from the cleavage site for Cas9 and 106 bp for Cas12a. A representative of two independent experiments is shown. d Representative Exo1-MR-pSae2-mediated resection of the plasmid-based DNA substrate treated with EcoRV, Cas9, or Cas12a, as indicated. DNA was stained with GelRed (Biotium). The DNA products, constituted mostly by mono- and dinucleotides, are not stained effectively by the dye. A representative of two independent experiments is shown. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41467-024-50080-y), licensed under a CC-BY license. Not internally tested by Novus Biologicals.