Anti-c-Myc antibody [Y69] (ab32072)

  Immunocytochemistry/ Immunofluorescence - c-Myc antibody [Y69] (ab32072)

ab32072 staining c-Myc in HEK-293 cells transfected with CACNB4-c-Myc by Immunocytochemistry/ Immunofluorescence.

Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100 then blocked using 5% serum for 20 minutes at 25°C. Samples were then incubated with ab32072 at a 1/250 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal, used at a 1/500 dilution.

Image courtesy of Dr Vladimir Milenkovic by Abreview.

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  Western blot - c-Myc antibody [Y69] (ab32072)

Anti-c-Myc antibody [Y69] (ab32072) at 1/10000 dilution + Jurkat cell lysate.

Predicted band size : 49 kDa
Observed band size : 57 kDa (why is the actual band size different from the predicted?)

  Immunocytochemistry/ Immunofluorescence - c-Myc antibody [Y69] (ab32072)

Ab32072, at a dilution of 1/250, staining Hela cells by Immunoflourescence.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - c-Myc antibody [Y69] (ab32072)

Ab32072, at a dilution of 1/50, staining c-Myc in paraffin embedded human skin carcinoma tissue by Immunohistochemistry.

  Immunocytochemistry/ Immunofluorescence - c-Myc antibody [Y69] (ab32072)

ICC/IF image of ab32072 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab32072, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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.