Blocking Solutions

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Tony Rook
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Blocking Solutions

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