Guide-itTM Cas9 Polyclonal Antibody

Description

The Guide-it Cas9 Polyclonal Antibody is a rabbit antibody raised against recombinant Cas9 protein fromStreptococcuspyogenes.This antibody recognizes three variants of the Cas9 nuclease: wild-type Cas9, Cas9 nickase, and Cas9 nuclease-deficient mutants. This antibody can recognize as little as 0.5 ng of recombinant Cas9 by Western blot. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back The CRISPR/Cas9 system, a simple, RNA-programmable method to mediate genome editing in mammalian cells.The CRISPR/Cas9 system relies on a single guide RNA sgRNA directing the Cas9 endonuclease to induce a double strand break at a specific target sequence three base-pairs upstream of a PAM sequence in genomic DNA. This DNA cleavage can be repaired in one of two ways: 1 nonhomologous end joining, NHEJ resulting in gene knockout due to error-prone repair orange , or 2 homology-directed repair HDR , resulting in gene knockin due to the presence of a homologous repair template purple . Back Back Back The Cas9 polyclonal antibody detects Cas9 protein by immunocytochemistry ICC .RPE cells stably expressing ZsGreen1 were transfected with 0.35 mug of a plasmid encoding wild-type Cas9 under the control of the SV40 promoter using Xfect Transfection Reagent. Cells were fixed with 4 % paraformaldehyde 48 hours post-transfection and stained with Cas9 polyclonal antibody 1:150 for one hour at room temperature RT , followed by staining with donkey anti-rabbit secondary antibody 1:100 for 30 min at RT. In the control, cells were only stained with the secondary antibody. Back Back