Paraffin embedded human breast carcinoma tissue stained for Estrogen Receptor beta using ab288 at 1/200 dilution in immunohistochemical analysis.

All lanes : Anti-Estrogen Receptor beta antibody [14C8] (ab288) at 1/5000 dilution

Lane 1 : Non-transfected HEK-293T whole cell extracts
Lane 2 : Transfected HEK-293T whole cell extracts

Lysates/proteins at 30 µg per lane.

Predicted band size: 59 kDa

7.5% SDS-PAGE

ab288 staining Estrogen Receptor beta in infiltrating lobular carcinoma of the human breast tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde. Endogenous peroxidase blocking was with 3% H₂O₂ at room temperature for 30 minutes. Blocking with 1.5% goat serum (dilute goat serum by 1xPBS), at room temperature for 30min; antigen retrieval was by heat mediation in a citrate buffer, pH 6.0, 15min (Cuisinart Electric Pressure Cooker #EPC-1200, choose "high pressure"). Samples were incubated with primary antibody (1/100) for overnight at 4°C. An ABC HRP Kit Mouse IgG) was used as the secondary antibody (1/200), room temperature for 30min. Washed twice for 5 minutes using PBS. DAB was used as a substrate.

All lanes : Anti-Estrogen Receptor beta antibody [14C8] (ab288) at 1/500 dilution

Lane 1 : Human brain tissue lysate
Lane 2 : Human liver tissue lysate
Lane 3 : Human intestine tissue lysate

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : Mouse IgG antibody (HRP) at 1/5000 dilution

Developed using the ECL technique.

Predicted band size: 59 kDa

7.5% gel.

Running condition: 80V, 15min; 140V, 40min.

Transfer condition: Semi-dry, 18 V, 60min (NC membrane)

blocking condition: 5% non-fat milk in TBST, RT, 60min.

primary antibody incubation: 4°C overnight.

Secondary antibody incubation: room temperature for 1 hour.

Washing conditions: 5 ml TBST, 4 x 5min.

ECL exposure. 

ab288 staining endocervical epithelium by IHC-P.