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.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|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.
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