---
title: Vina's Buffer Library
description: The complete library of buffers!
---

[Blog ](https://www.miss-academia.com/blog)

# [Vina's Buffer Library](https://www.miss-academia.com/blog/vinas-buffer-library)

 Written by [Vina Putra](https://www.miss-academia.com/blog/author/vina-putra) | Jul 17, 2020 1:20:24 PM

#### Check out my ever growing buffer library to help you prepare your own buffers!

| ##### **General buffers** |
| --- |
| ** ** |
| **1 X PBS:** |
| To make 1 L |
| 8 g NaCl (mw: 58.4 g/mol) |
| 200 mg KCl (mw: 74.551 g/mol) |
| 1.44 g Na2HPO4 (mw: 141.96 g/mol) |
| 245 mg KH2PO4 (mw: 136.086 g/mol) |
| Add distilled water to make volume 1 L |
| Adjust pH to 7.4 |
|   |
| **0.5 M EDTA, pH 8:** |
| Add 400 mL distilled water |
| Add 93.05 g Na2•EDTA•2H2O (disodium dyhydrate) |
| 9 g solid NaOH |
| Adjust the pH with with 10 N NaOH |
| Add distilled water to make volume 500 mL |
|   |
| **10 M NaOH (= 10 N NaOH): ** |
| To make 500 mL |
| 200 g NaOH in a final volume of 500 mL distilled water |
|   |
| **5 M NaCl:** |
|  To make 1 L  |
| 292.2 g NaCl in a final volume of 1 L distilled water |
|   |
| **1 M HEPES stocks: ** |
| To make 1 L |
| 238.3 g HEPES (free acid)  |
| 800 mL of ddH2O  |
| Adjust the pH to the desired value with 10 N NaOH |
| Add distilled water to make volume 1 L |

| ##### **For SDS PAGE** |
| --- |
|   |
| **10x Tris-glycine running buffer:** |
| To make 4 L |
| 121.1 g Tris base |
| 576 g glycine |
| 200 mL 20% SDS |
| Add distilled water to make volume 4 L |
|   |
| **1 x Running buffer:** |
| To make 1 L |
| 100 mL 10x running buffer |
| 900 ml distilled water |
|   |
| **For making main separating gels:** |
| 1.5 M Tris, pH 8.8 : room temperature |
| To make 1 L |
| 800 mL distilled water |
| 181.65 g Tris base |
| Adjust the pH to 8.8 with concentrated HCl |
| Add distilled water to make volume 1 L |
|   |
| **For making stacking gels:** |
| 1.5 M Tris, pH 6.8 |
| 181.65 g Tris base |
| Adjust the pH to 6.8 with concentrated HCl |
| Add distilled water to make volume 1 L |
|   |

 

 

| ##### **For western blot** |
| --- |
| ** ** |
| **10x Transfer buffer:** |
| To make 4 L |
| 121.1 g Tris base |
| 576 g glycine |
| Bring up the volume to 4 L with ddH2O |
|   |
| **1x Transfer buffer: 4 C temperature** |
| To make 1 L |
| 700 mL cold ddH2O |
| 100 mL 10x Transfer buffer |
| 200 mL methanol |
|   |
| **Blocking buffer: 1-3% usually works** |
| to make 100 ml |
| 1-3 grams of skim milk powder |
| add 1X PBS to make volume 100 ml |
|   |
| **20x TBS:** |
| To make 4 L |
| 193.6 g Tris base |
| 640 g NaCl |
| Add distilled water to make volume 3.2 L |
| Adjust pH to 7.6 by adding HCl |
| Add distilled water to make volume 4 L |
|   |
| **20X TBST:** |
| To make 100 mL |
| 2 mL Tween-20 |
| 100 mL of 20x TBS |
|   |
| **1X TBST: also works using 1X PBS** |
| To make 1 L |
| Add 500 ul Tween-20 |
| Add 1X PBS to make volume to 1 L |

 

|   |
| --- |
| **RIPA buffer:** |
| To make 10 ml |
| NaCl (1 M) 1.5 ml to make cocentration 150 mM |
| Nonidet P-40 (1%) 0.1 ml in 10 ml |
| Sodium deoxycholate  (DOC) (0.5%) 0.05 ml |
| SDS (0.1%) 0.01 mL |
| Tris (50 mM, pH 7.4) 5 ml to make concentration 25 mM |
| Add distilled water to make total volume 10ml |
|   |

| **6X Loading buffer** |
| --- |
| 0.3 M Tris-HCl |
| 0.6 M DTT |
| 10% SDS |
| 0.06% Bromophenol blue |
| 30% Glycerol |

 

 

 

[View full post](https://www.miss-academia.com/blog/vinas-buffer-library)

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