All lanes : Anti-HuR / ELAVL1 antibody [19F12AE12] (ab170193) at 1 µg/mlLane 1 : H9C2 Rat Cell LysateLane 2 : H4IIE Rat Cell LysateLane 3 : MEF Mouse Cell LysateLane 4 : 3T3 Mouse Cell LysateLane 5 : SH-SY5Y Cell LysateLane 6 : HepG2 Cell LysateLysates/proteins at 20 µg per lane.SecondaryGoat anti-mouse AP at 1/5000 dilutiondeveloped using the ECL technique

Immunocytochemistry on HeLa cells.Cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15minutes). The cells were then incubated with the anti-HuR antibody (5µg/mL) for 2 hours at room temperature or over night at 4°C. The secondary antibody used was (red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1 hour. A cytoplasmic antibody (ab75834) was used as counter stain (green). 10% goat serum was used as a blocking agent for all blocking steps.The target protein localizes mainly to the nucleus.

Immunocytochemistry on SH-SY5Y cells (Human neuroblastoma cell line).Cells cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were then incubated with the anti-HuR antibody (5µg/mL) for 2 hours at room temperature or overnight at 4°C. The secondary antibody was (red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. A cytoplasmic antibody (ab75834) was used as counter stain (Green). 10% Goat serum was used as a blocking agent for all blocking steps.The target protein localizes mainly to the nucleus.  

Flow cytometric analysis of methanol-fixed HeLa cells labeling HuR / ELAVL1 with ab170193 at 1µg/mL (blue) or an equal amount of an isotype control antibody (red).