TLR9, Mouse, mAb 5G5 Antibody

For immunohistology, flow cytometry and Western blotting, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.

F: 10 μm 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. (Ref.7) IF: cells were fixed with 2% formalin for 15 minutes at RT and permeabilized with a mAb (4μg/400μl) containing buffer (PBS, 0.2% BSA, 0.2% saponin) - for 1 hour. (Ref.1) FC: RAW264.7 cells were fixed for 15 minutes with 4% formalin and permeabilized (PBS, 0.5%BSA, 0,5% saponin) at RT. (Ref.1) P: paraffin embedded tissues 5μm 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 37°C. (Ref.3) W: reduced lysates were resolved by 10% SDS-PAGE and blotted on nitrocellulose. After blocking with 5% skimmed milk TLR9 was detected with 2 μg/ml 5G5. (Ref.1)