Synthesized non-phosphopeptide derived from human Src around the phosphorylation site of tyrosine 529.
Our Abpromise guarantee covers the use of ab47405 in the following tested applications.
|ICC/IF||Use a concentration of 1 - 5 µg/ml.|
|WB||1/500 - 1/1000. Detects a band of approximately 60 kDa (predicted molecular weight: 60 kDa).|
Peptide ELISA only.
|IHC-P||Use at an assay dependent concentration.|
ab47405 staining Src in Pig retinal pigment epithelium primary cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilzed with 0.5% Triton X-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/1000 in 0.1% TX100, 1% goat serum, 1XPBS) for 16 hours at 4°C. An Alexa Fluor®488-conjugated goat anti-rabbit IgG polyclonal (1/5000) was used as the secondary antibody. Nuclei were counterstained with DAPI.
ICC/IF image of ab47405 stained MCF7 cells (ab3871). The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47405, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
This image is courtesy of an anonymous abreview.Primary antibody incubated for 1 hour at 25°C.Blocked with 3& BSA for 1 hour at 25°C.Gel run under denaturing conditions.Detection method: ImmunoStar.