ab64116 (1:40) staining CD80 in paraffin-embedded mouse lymph node tissue (left panel) using an automated system (Ventana Discovery). Right-hand panel shows negative control (no primary antibody). Using this protocol there is strong membrane staining, of activated B cells in the germinal centres. What appear to be scattered activated B cells or possibly plasma cells in the interfollicular areas of the lymph node and blood vessels.
Sections were rehydrated and antigen retrieved in CC1 Cell Conditioning Buffer using Ventana Extended Retrieval programme. Slides were blocked in 3% H2O2 / 4 min / 37°C and incubated with ab64116 (1:40 dilution / 1 hour / 37°C). Sections then blocked (4mins / 37°C) and incubated with Dako swine anti-rabbit antibody (1:50, 28 min / 37°C). Staining was amplified and detected by incubation with Ventana Streptavidin ABC system (16 min / 37°C) and Ventana DAB map reagent (8
Western blot - CD80 antibody (ab64116)
All lanes : Anti-CD80 antibody (ab64116) at 1 µg/ml
Lane 1 : Spleen (Mouse) Tissue Lysate Lane 2 : Thymus (Mouse) Tissue Lysate Lane 3 : Brain (Mouse) Tissue Lysate Lane 4 : Skeletal Muscle (Mouse) Tissue Lysate Lane 5 : Spinal Cord (Mouse) Tissue Lysate Lane 6 : EL4 whole cell lysate (ab7183) Lane 7 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution Developed using the ECL technique
CD80 has a predicted molecular weight of 33 kDa, however it is predicted to be extensively glycosylated (SwissProt data). The band seen at 65 kDa is consistent with the banding pattern observed for other commercially available antibodies to CD80. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.