Application: |
Immunohistochemistry |
Cells used: |
Brain perfused fixed brain tissue |
Brief Protocol: |
AD Tg rats were perfused with 0.9% heparnized saline (100ml) followed by 10% neutral buffered formalin (150ml). Then the brains were post fixed in 10% neutral buffered formalin+ 20% sucrose overnight at 4C. Brains were frozen in dry ice, cut in Leica microtome at 25um thickness. The brain sections were incubated in 0.5% H2O2 for 15 min, followed by PBS containing Triton X for 30 min and then incubated in 5% normal goat serum for 20 min. Then the sections were incubated in 4G8 antibody at 1:500 and 1:1K for overnight at 4C, followed by biotinylated secondary IgG at 1:200 dilution and then incubated in ABC solution (Vector labs; 1:100) for 1 hour. Finally sections were stained with combination of H2O2+ DAB-HCl. The reaction product appears brown. Sections were mounted onto charged slides. Dried in slide warmer and cleared in Xylene and coverslipped with DPX mountant. |
Results Summary: |
The 4G8 antibody labelled all the plaques in the brain, mostlty in the parenchyma but few vascular plauqes were also labeled. |
Additional Notes: |
This is an excellent antibody to identify amyloid abeta fragment 17-24 in the brain. |
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