TLR9 Antibody (5G5)

IHC-Frozen: 10um sections were fixed with acetone for 10 minutes. PBS washed sections were incubated with 5G5 1:100 in 1% BSA for 30 minutes at RT. IF: cells were fixed with 2% formalin for 15 minutes at RT and permeabilized with a mAb (4ug/400ul) containing buffer (PBS, 0.2% BSA, 0.2% saponin) for 1 hour. FC: RAW264.7 cells were fixed for 15 minutes with 4% formalin and permeabilized (PBS, 0.5%BSA, 0,5% saponin) at RT. IHC-Paraffin: paraffin embedded tissues 5um sections were made. After antigen retrieval (0.01mol/l, pH6 sodium citrate) and quenching of endogenous peroxidase, sections were blocked with 0.5% ovalbumin and 0.1% gelatin for 20 minutes at RT. Sections were incubated with 5G5 for 1 hour at 37C. WB: reduced lysates were resolved by 10% SDS-PAGE and blotted on nitrocellulose. After blocking with 5% skimmed milk TLR9 was detected with 2 ug/ml 5G5