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Formalin-fixed and paraffin-embedded human testis tissue reacted with ab75483 at a dilution of 1/50. A peroxidase-conjugated secondary antibody was then used, followed by DAB staining.
All lanes : Anti-Lin28 antibody [55CT58.12.1] (ab75483) at 1 µg/ml
Lane 1 :
Lane 2 :
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (ab97040) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 23 kDa
Observed band size : 28 kDa (why is the actual band size different from the predicted?)
Additional bands at : 72 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 12 minutes
The predicted molecular weight of Lin28 is 23 kDa (SwissProt), however we expect to observe a banding pattern at 28 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
ICC/IF image of ab75483 stained MES cells. The cells were 4% formaldehyde 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 (ab75483, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) 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.
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