SM buffer with gelatin is a commonly used buffer solution for various cellular and biochemical applications. It is a versatile buffer that can be used for cell lysis, protein extraction, and as a stabilizing agent for cell suspensions. The preparation of SM buffer with gelatin involves mixing the following components in distilled water:
Ingredients:
- NaCl (150 mM)
- MgSO4 (10 mM)
- KCl (5 mM)
- Na2HPO4 (10 mM)
- KH2PO4 (10 mM)
- Gelatin (0.5%)
Procedure:
- Dissolve 150 mM NaCl, 10 mM MgSO4, 5 mM KCl, 10 mM Na2HPO4, and 10 mM KH2PO4 in distilled water.
- Adjust the pH to 7.4 using NaOH or HCl.
- Sterilize the solution by filtration or autoclaving.
- Add 0.5% gelatin to the solution and mix until dissolved.
- Store the SM buffer with gelatin at 4°C.
Note: Gelatin should be added just before use, as it can cause the buffer to become turbid over time. Also, the concentration of gelatin can be adjusted depending on the specific application and requirements.
To prepare one liter follow this
To prepare 1 liter of SM buffer with gelatin, dissolve the NaCl and MgSO4·7H2O in 800 ml of H2O; add the Tris-Cl and gelatin; and adjust the volume to 1 liter with H2O. Sterilize the buffer by autoclaving for 20 minutes at 15 psi (1.05 kg/cm2) on a liquid cycle. After the solution has cooled, dispense 50 ml aliquots into sterile containers. SM buffer with gelatin may be stored indefinitely at room temperature. Discard each aliquot after use to minimize the chance of contamination. To prepare SM Buffer without gelatin click here and to prepare Tris-cl click here
| Reagent | Amount | Final concentration |
|---|---|---|
| NaCl | 5.8 g | 100 mM |
| MgSO4·7H2O | 2 g | 8 mM |
| Tris-Cl (1 M, pH 7.5) | 50 ml | 50 mM |
| Gelatin (2%, w/v) | 5 ml | 0.01% (w/v) |
| H2O | to 1 liter |
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