ImmunoHistochemistry Buffer Book

10/26/2018 IHC Buffer Recipes | Thermo Fisher Scientific - IN
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IHC Buffer Recipes
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Buffer 1: 10 mM PBS, pH 7.4
Buffer 2: 0.1 M NaHCO3, pH 9.6
Buffer 3: 10 m M PBS, pH 7.4 with TWEEN 20
Buffer 4: 0.1 M Citrate, pH 4.2 with 0.03% H2O2
Buffer 5: 50 mM Tris-buffered Saline, pH 7.5
Buffer 6: 0.1 M Amino-Methyl- Propanediol, pH 10.3
Buffer 7: 0.1 M Citrate-Phosphate, pH 5.0, with 0.03% H2O2
Buffer 8: 0.1 M Diethanolamine, pH 10.3
Buffer 9: 20 mM Sodium Phosphate, pH 7.0
Buffer 10: 50 mM Tris-buffered Saline, pH 7.4
Buffer 11: 20 mM Tris-buffered Saline, pH 8.2 with 0.1% BSA
Buffer 12: 10 mM PBS, pH 8.0 with 0.1% BSA
Buffer 13: 0.2 M Sodium Citrate, pH 3.5
Buffer 14: 10 mM Sodium phosphate saline, pH 7.0
Buffer 15: 100 mM PBS, pH 7.4, with 0.01% Proclin
Buffer 16: 0.1 M Glycine-HCl, pH 2.7
Buffer 17: 0.1 M Acetic acid, pH 2.8
Recipe for Buffer 1: 10 mM PBS, pH 7.4

Note: These recipes are designed to make the common buffers mentioned in our procedures. This list is not all inclusive. Use NaOH or HCl to adjust pH,
being careful not to overshoot and back-titrate, as this may alter salt concentration more than necessary.
Add 1 bag of PBS (Cat. No. 00-3000) to 1L of dH20
or combine:
0.26 g KH2PO4
2.17 g Na2HPO4-7H20
8.71 g NaCl
800 ml dH20
Adjust pH to 7.4 and bring volume to 1 L with dH20
Recipe for Buffer 2: 0.1 M NaHCO 3 , pH 9.6

Combine:
8.4 g NaHCO3
800 ml dH20
Adjust pH to 9.6 and bring volume to 1 L with dH20.
Recipe for Buffer 3: 10 m M PBS, pH 7.4 with TWEEN 20

Prepare 1L of Buffer 1 , then add 0.5 mL of TWEEN 20. Mix Well
or combine:
0.26 g KH2PO4
2.17 g Na2HPO4-7H20
https://www.thermofisher.com/in/en/home/life-science/cell-analysis/cellular-imaging/ihc/zymed-buffer-recipes.html 1/4
8.71 g NaCl
800 ml dH20
Adjust pH to 7.4 and bring volume to 1 L with dH20.
Recipe for Buffer 4: 0.1 M Citrate, pH 4.2 with 0.03% H 2 O 2

Combine:
9.802 g citric acid
14.4 g sodium citrate-2H20
Adjust pH to 4.1
Add 1 mL 30% H2O2
Bring volume to 1L with dH20.
Recipe for Buffer 5: 50 mM Tris-buffered Saline, pH 7.5

Combine:
6.35 g Tris-HCl
1.18 g Tris Base
8.77 g NaCl
800 ml dH2O
Adjust pH to 7.5 and bring volume to 1 L with dH2O.
Recipe for Buffer 6: 0.1 M Amino-Methyl- Propanediol, pH 10.3

Combine:
10.5 g AMPD [HOCH2C(CH3)(NH2)CH2OH]
800 mL dH2O
Recipe for Buffer 7: 0.1 M Citrate-Phosphate, pH 5.0, with 0.03% H 2 O 2

Combine:
4.7 g Citric Acid
9.2 g Dibasic Sodium Phosphate (Na2HPO4) dehydrate
Recipe for Buffer 8: 0.1 M Diethanolamine, pH 10.3

Combine:
10.5 g (HOCH2CH2)2NH
800 mL dH2O
Recipe for Buffer 9: 20 mM Sodium Phosphate, pH 7.0

Combine:
3.27 g Na2HPO4-7H2O
0.94 g NaH2PO4
Recipe for Buffer 10: 50 mM Tris-buffered Saline, pH 7.4

Combine:
6.61 g Tris-HCl
0.97 g Tris Base
8.77 g NaCl
800 ml dH2O
Recipe for Buffer 11: 20 mM Tris-buffered Saline, pH 8.2 with 0.1% BSA
Combine:
2.42 g Tris Base
1.3 g Sodium azide
8.77 g NaCl
1 g Bovine Serum Albumin
800 ml dH2O
Recipe for Buffer 12: 10 mM PBS, pH 8.0 with 0.1% BSA

Combine:
0.43 g KH2PO4
2.79 g Na2HPO4-7H2O
8.77 g NaCl
1.3 g Sodium azide
1 g Bovine serum albumin
800 ml dH2O
Adjust pH to 8.0 and bring volume to 1 L with dH2O
Recipe for Buffer 13: 0.2 M Sodium Citrate, pH 3.5

Combine:
25 g Sodium citrate-2H2O
40 g Citric acid
7.2 g NaCl
800 ml dH2O
Recipe for Buffer 14: 10 mM Sodium phosphate saline, pH 7.0

Combine:
1.64 g Na2HPO4-7H2O
0.47 g NaH2PO4
8.77 g NaCl
Recipe for Buffer 15: 100 mM PBS, pH 7.4, with 0.01% Proclin
Combine:
2.6 g KH2PO4
21.7 g Na2HPO4-7H2O
8.77 g NaCl
1 ml proclin
800 ml dH2O
Recipe for Buffer 16: 0.1 M Glycine-HCl, pH 2.7

Combine:
11.1 g Glycine-HCl
800 ml dH2O
Recipe for Buffer 17: 0.1 M Acetic acid, pH 2.8

Combine:
5.72 mL Glacial Acetic Acid
800 ml dH2O
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ImmunoHistochemistry Buffer Book

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10/26/2018 IHC Buffer Recipes | Thermo Fisher Scientific - IN

IHC Buffer Recipes

Buffer 1: 10 mM PBS, pH 7.4

Buffer 2: 0.1 M NaHCO3, pH 9.6

Buffer 3: 10 m M PBS, pH 7.4 with TWEEN 20

Buffer 4: 0.1 M Citrate, pH 4.2 with 0.03% H2O2

Buffer 5: 50 mM Tris-buffered Saline, pH 7.5

Buffer 6: 0.1 M Amino-Methyl- Propanediol, pH 10.3

Buffer 7: 0.1 M Citrate-Phosphate, pH 5.0, with 0.03% H2O2

Buffer 8: 0.1 M Diethanolamine, pH 10.3

Buffer 9: 20 mM Sodium Phosphate, pH 7.0

Buffer 10: 50 mM Tris-buffered Saline, pH 7.4

Buffer 11: 20 mM Tris-buffered Saline, pH 8.2 with 0.1% BSA

Buffer 12: 10 mM PBS, pH 8.0 with 0.1% BSA

Buffer 13: 0.2 M Sodium Citrate, pH 3.5

Buffer 14: 10 mM Sodium phosphate saline, pH 7.0

Buffer 15: 100 mM PBS, pH 7.4, with 0.01% Proclin

Buffer 16: 0.1 M Glycine-HCl, pH 2.7

Buffer 17: 0.1 M Acetic acid, pH 2.8

Recipe for Buffer 1: 10 mM PBS, pH 7.4

Add 1 bag of PBS (Cat. No. 00-3000) to 1L of dH20

Adjust pH to 7.4 and bring volume to 1 L with dH20

Recipe for Buffer 2: 0.1 M NaHCO 3 , pH 9.6

Adjust pH to 9.6 and bring volume to 1 L with dH20.

Recipe for Buffer 3: 10 m M PBS, pH 7.4 with TWEEN 20

Prepare 1L of Buffer 1 , then add 0.5 mL of TWEEN 20. Mix Well

Adjust pH to 7.4 and bring volume to 1 L with dH20.

Recipe for Buffer 4: 0.1 M Citrate, pH 4.2 with 0.03% H 2 O 2

9.802 g citric acid

14.4 g sodium citrate-2H20

Add 1 mL 30% H2O2

Bring volume to 1L with dH20.

Recipe for Buffer 5: 50 mM Tris-buffered Saline, pH 7.5

1.18 g Tris Base

Adjust pH to 7.5 and bring volume to 1 L with dH2O.

Recipe for Buffer 6: 0.1 M Amino-Methyl- Propanediol, pH 10.3

10.5 g AMPD [HOCH2C(CH3)(NH2)CH2OH]

Adjust pH to 10.3 and bring volume to 1 L with dH2O.

Recipe for Buffer 7: 0.1 M Citrate-Phosphate, pH 5.0, with 0.03% H 2 O 2

4.7 g Citric Acid

9.2 g Dibasic Sodium Phosphate (Na2HPO4) dehydrate

Adjust pH to 5.0 and bring volume to 1 L with dH2O.

Recipe for Buffer 8: 0.1 M Diethanolamine, pH 10.3

Adjust pH to 10.3 and bring volume to 1 L with dH2O.

Recipe for Buffer 9: 20 mM Sodium Phosphate, pH 7.0

Adjust pH to 7.0 and bring volume to 1 L with dH2O.

Recipe for Buffer 10: 50 mM Tris-buffered Saline, pH 7.4

0.97 g Tris Base

Adjust pH to 7.4 and bring volume to 1 L with dH2O.

2.42 g Tris Base

1.3 g Sodium azide

1 g Bovine Serum Albumin

Adjust pH to 8.2 and bring volume to 1 L with dH2O.

Recipe for Buffer 12: 10 mM PBS, pH 8.0 with 0.1% BSA

1.3 g Sodium azide

1 g Bovine serum albumin

Adjust pH to 8.0 and bring volume to 1 L with dH2O

Recipe for Buffer 13: 0.2 M Sodium Citrate, pH 3.5

Adjust pH to 3.5 and bring volume to 1 L with dH2O

Recipe for Buffer 14: 10 mM Sodium phosphate saline, pH 7.0

Adjust pH to 7.0 and bring volume to 1 L with dH2O.

Adjust pH to 7.4 and bring volume to 1 L with dH2O.

Recipe for Buffer 16: 0.1 M Glycine-HCl, pH 2.7

Adjust pH to 2.7 and bring volume to 1 L with dH2O.