Mon-Fri 9:00-5:00
IHC Washing Buffer- Tris Buffered Saline (TBS)
Home » Protocols  »  IHC Washing Buffer Protocols  »  IHC Washing Buffer- Tris Buffered Saline (TBS)
IHC Washing Buffer- Tris Buffered Saline (TBS)

10X TBS (0.5M Tris Base, 9% NaCl, pH 8.4):

       Trizma base ------------------------------- 61 g

       NaCl ---------------------------------------- 90 g

       Distilled water ---------------------------- 1000 ml

       Mix to dissolve and adjust pH to 8.4 using concentrated HCl.

       Store this solution at room temperature. Dilute 1:10 with distilled water before use and adjust pH if necessary.

20X TBS (1M Tris Base, 18% NaCl, pH 8.4):

       Trizma base -------------------------------- 122 g

       NaCl ----------------------------------------- 180 g

       Distilled water ----------------------------- 1000 ml

       Mix to dissolve and adjust pH to 8.4 using concentrated HCl.

  Store this solution at room temperature. Dilute 1:20 with distilled water before use and adjust pH if necessary.

10X TBS-Tween 20 (0.5M Tris Base, 9% NaCl, 0.5% Tween 20, pH 8.4):

       Trizma base -------------------------------- 61 g

       NaCl ----------------------------------------- 90 g

       Distilled water ----------------------------- 1000 ml

       Mix to dissolve and adjust pH to 8.4 using concentrated HCl and then add 5 ml Tween 20.

       Store this solution at room temperature. Dilute 1:10 with distilled water before use and adjust pH if necessary.

20X TBS-Tween 20 (1M Tris Base, 18% NaCl, 1% Tween 20, pH 8.4):

       Trizma base -------------------------------- 122 g

       NaCl ----------------------------------------- 180 g

       Distilled water ----------------------------- 1000 ml

       Mix to dissolve and adjust pH to 8.4 using concentrated HCl and then add 10 ml Tween 20.

       Store this solution at room temperature. Dilute 1:20 with distilled water before use and adjust pH if necessary.

Note: TBS is used for immunohistochemical staining when background is high or when alkaline phosphatase conjugated antibodies are used. TBS is often used for diluting secondary antibodies or streptavidin-AP conjugate. TBS-Tween 20 is often used for washing steps.

Leave a Reply