Please find the following media preparation methods for:
Blocking Solutions
Peroxidase Blocking Soluiton (3% H2O2 in PBS):
30% H2O2 -------------------------- 2 ml
1X PBS ----------------------------- 18 ml
Mix well and store at 4 C for up to 3 months.
Block sections for 10 minutes after primary antibody
incubation. Note: this solution is recommended for paraffin
sections.
Peroxidase Blocking Soluiton (0.3% H2O2 in Methanol):
30% H2O2 ------------------------- 0.2 ml
Methanol -------------------------- 18 ml
Mix well and store at 4 C.
Block sections for 20-30 minutes after primary antibody
incubation. Note: this solution is recommended for frozen
sections.
Avidin/Biotin Block:
Avidin 0.001% in PBS
Biotin 0.001% in PBS
Store these blocking solution at 4 C.
Incubate sections for 10-15 min each and rinse with PBS between steps. Recommended to block before primary antibody incubation.
Normal Goat Serum Blocking Solution:
2% goat serum (blocking)
1%BSA (stabilizer)
0.1% cold fish skin gelatin (blocking)
0.1% Triton X-100 (penetration enhancer)
0.05% Tween 20 (detergent and surface tension reducer)
0.05% sodium azide (preservative)
0.01M PBS, pH 7.2
Mix well and store at 4 C.
Normal Horse Serum Blocking Solution:
2% horse serum (blocking)
1%BSA (stabilizer)
0.1% cold fish skin gelatin (blocking)
0.1% Triton X-100 (penetration enhancer)
0.05% Tween 20 (detergent and surface tension reducer)
0.05% sodium azide (preservative)
0.01M PBS, pH 7.2
Mix and store at 4 C.
Normal Swine Serum Block Solution:
2% swine serum (blocking)
1%BSA (stabilizer)
0.1% cold fish skin gelatin (blocking)
0.1% Triton X-100 (penetration enhancer)
0.05% Tween 20 (detergent and surface tension reducer)
0.05% sodium azide (preservative)
0.01M PBS, pH 7.2
Mix and store at 4 C.
Normal Donkey Serum Block Solution:
2% donkey serum (blocking)
1%BSA (stabilizer)
0.1% cold fish skin gelatin (blocking)
0.1% Triton X-100 (penetration enhancer)
0.05% Tween 20 (detergent and surface tension reducer)
0.05% sodium azide (preservative)
0.01M PBS, pH 7.2
Mix and store at 4 C.
Normal Rabbit Serum Block Solution:
2% rabbit serum (blocking)
1%BSA (stabilizer)
0.1% cold fish skin gelatin (blocking)
0.1% Triton X-100 (penetration enhancer)
0.05% Tween 20 (detergent and surface tension reducer)
0.05% sodium azide (preservative)
0.01M PBS, pH 7.2
Mix and store at 4 C.
Link Reference:
http://www.ihcworld.com/_protocols/blocking_solutions/blocking_solutions.htm