Anti-Eph receptor A4/SEK 抗体 (ab5389)

Blocking/Dilution buffer: 5% NFDM/TBST.

ab5389 EphA4 antibody immunostaining cortical neurons (image taken at X2; cytoplasmic staining).Bar=20 microns.

Protocol: ab5389 incubated overnight at room temperature at a dilution of 1/100-1/300 on free floating perfusion fixed coronal sections of brain. Immunostaining visualised by direct fluorescence (Alexa Fluor^® 488, 1/1000).

ab5389 EHpA4 antibody immunostaining hippocampal neurons ( X2 magnification; cytoplasmic staining). Bar: 20 microns.

Protocol: ab5389 incubated overnight at room temperature at a dilution of 1/100-1/300 on free floating perfusion fixed coronal sections of brain. Immunostaining visualised by direct fluorescence (Alexa Fluor^® 488, 1/1000).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma (BC) tissue labeling Eph receptor A4/SEK with ab5389 at 1/25. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at 37°C. Heat mediated antigen retrieval was performed using a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hour at 37°C. A Peroxidase-conjugated Goat anti-rabbit polyclonal (ready to use) was used as the secondary antibody.

Flow cytometry analysis of CEM cells labelling Eph receptor A4/SEK (green) with ab5389 compared to a negative control (blue). The cells were fixed with paraformaldehyde and then permeabilized with 90% methanol for 10 min. The cells were then incubated in 3% BSA to block non-specific protein-protein interactions followed by the antibody (ab5389, 1µg/1x10⁶ cells) for 60 min at 37ºC. The secondary antibody used was FITC conjugated Goat anti-Rabbit antibodies at 1/200 dilution for 40 min at room temperature. Blank control (blue line) was used under the same conditions. Acquisition of >10,000 events was performed.