Cas9 Monoclonal Antibody
- High specificity: very low background on mammalian cell lysates
- High sensitivity: detect less than 1 ng of Cas9
The Guide-it Cas9 Monoclonal Antibody (Clone TG8C1) is a mouse antibody produced by hybridoma cells against full-length recombinant Cas9 protein from Streptococcus pyogenes. This antibody can recognize less than 1 ng of wild-type Cas9 in bacterial and mammalian cell lysates by Western blot.
Cas9 Monoclonal Antibody Sensitivity on Western blot
The Guide-it Cas9 Monoclonal Antibody (Clone TG8C1) was tested in Western blot analysis of either rCas9 (Panel A) or lysates made from HEK 293 cells transiently expressing the Cas9 protein after plasmid transfection (Panel B). The Western blot was probed with a 1:3,000 dilution of the antibody, and detection was performed using an anti-mouse HRP-coupled secondary antibody at a 1:10,000 dilution.
Panel A: Increasing amounts of rCas9 showed clearly visible band at the expected molecular weight of ~160 kDa, even for the smallest amount loaded (1 ng).
Panel B: A clearly visible Cas9 band at ~160 kDa was detected only in the lanes of transfected cells. As few as 100 cells were sufficient for detection.
Immunocytochemistry with Cas9 Monoclonal Antibody
RPE cells stably expressing ZsGreen1 were transfected with plasmid encoding wild-type Cas9 using Xfect Transfection Reagent. Cells were fixed with 4% paraformaldehyde 48 hours post-transfection, and stained with Cas9 monoclonal antibody (1/100) for one hour at room temperature (RT), followed by staining with donkey anti-mouse secondary antibody (1/1000). The antifade mounting media contained DAPI, allowing for nuclear staining.