Buffer Recipes2021-02-18T16:16:49-05:00

To use the Visikol® HISTO-M™ approach with immunolabeling, several buffers are required which can either be purchased from Visikol through the Visikol store or can be made using common laboratory reagents.

Recipes for 100 mL – prepare in volumetric flask

Buffer nameCompositionRecipe for 100 mLStorage
PBST· PBS (1X)
· Tween® 20 (0.2% wt.)*
· 200 mg Tween® 20
· Dilute to 100 mL with 1X PBS
RT
Penetration Buffer· PBS (1X)
· 0.2% Tween® 20*
· 0.3M Glycine
· 20% DMSO
· 2.25 g Glycine
· Add 20 mL 1X PBST
· 20 mL DMSO
· Dilute to 100mL with ~60 mL PBST
RT
10X PTwH (10X washing buffer)· 10X PBS
· 2% Tween® 20
· 100 ug/mL Heparin
· 10 mg Heparin
· Add 20 mL 10X PBS
· 2g Tween 20
· Dilute to 100 mL with 10X PBS
4°C, 6 months.
Add 0.05% sodium azide
for long term storage
Blocking buffer· PBS
· 0.2% Tween 20*
· 6% Donkey Serum
· 10% DMSO
· 6 mL Donkey Serum
· Add 10 mL PBST
· Add 10 mL DMSO
· Dilute to 100 mL with PBST
4° C
Add 0.05% sodium azide
for long term storage
Antibody buffer· PBS
· 0.2% Tween® 20*
· Heparin
· 3% Donkey Serum
· 5% DMSO
· 3 mL Donkey Serum
· Add 20 mL 1X Washing Buffer
· Add 5 mL DMSO
· Dilute to 100 mL with 1X Washing Buffer
4° C
Add 0.05% sodium azide
for long term storage
  • To make 100 mL of 1X washing buffer, dilute 10 mL 10X buffer with 90 mL of DI H2O
  • Sodium azide will interfere with horseradish peroxidase-based protocols, but has no effect on immunofluorescent labeling

Catalog numbers

  1. Tween® 20– Fisher Scientific – Catalog # BP337
  2. Glycine – Fisher Scientific – Catalog # G46
  3. DMSO – Fisher Scientific – Catalog # BP231
  4. Heparin – Alfa Aesar – Catalog # A16198
  5. Normal Donkey Serum – Jackson Immunoresearch – Catalog #: 017-000-121

*Tween is a registered trademark of Croda International PLC.

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