| 1. Pretreatment with Hyaluronidase
(Negative control) |
| |
Treat with reaction buffer of Hyaluronidase
(100mM Sodium acetate buffer, pH6.0) for 15min at 37°C. |
| |
 No wash |
| |
Treat with Hyaluronidase (200TRU/mL; 100mM Sodium
acetate buffer, pH6.0) for 2hrs at 60°C. |
| |
Wash
with PBS |
| 2. Blocking endogeneous avidin
biotin activity |
| |
Treat with avidin solution for 20min at RT. |
| |
Wash |
| |
Treat with biotin solution for 20min at RT. |
| |
Wash |
| 3. Treat with 0.1% BSA solution
for 1hr at RT. |
| |
Wash |
| 4. Treat with Biotinylated HABP
(2µg/mL) for 1-2hrs at RT. |
| |
Wash |
| 5. Treat with Fluorophore conjugated
Streptavidin for 15min at RT. |
Histochemistry for Hyaluronic acid using HABP
