Anti-P2RX7 antibody (ab77413)

  Immunocytochemistry/ Immunofluorescence - P2RX7 antibody (ab77413)

ICC/IF image of ab77413 stained HeLa cells. 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 (ab77413, used at a 1/100 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) 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.

  Western blot - P2RX7 antibody (ab77413)

All lanes : Anti-P2RX7 antibody (ab77413) at 1/250 dilution

Lane 1 : U937 (Human leukemic monocyte lymphoma cell line) Whole Cell Lysate
Lane 2 : Thymus (Human) Tissue Lysate - adult normal tissue (ab30146)

Lysates/proteins at 10 µg per lane.

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 68,5 kDa
Observed band size : 74 kDa (why is the actual band size different from the predicted?)
Additional bands at : 45 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 1 minute

P2RX7 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.