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Blotting Resources
Download the
full blotting apparatus catalog
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full blotting apparatus catalog
pdf
Recommended Voltages
Common Buffers and their Recipe's
Additional Notes
TechnoSource range of Electro blotting apparatus
are manufactured in India.
| Instrument | Voltage for overnight transfer | Voltage for 1 hr transfer |
| Microblot | 25V | 80V |
| Microblot-T | 25V | 80V |
| Midiblot | 35V | 90V |
| Midiblot-T | 35V | 90V |
| Technoblot-O | 40V | 100V |
| Technoblot | 40V | 110V |
| Technoblot-F | 40V | 120V |
Common Buffers and their Recipe's
| Towbin buffer, 1L, 10x 25mM Tris, 192 mM glycine, 20% (v/v) methonol (pH 8.3) | |
| Tris base | 3.03 g |
| Glycine | 14.4 g |
| D/w | 500 mL |
| Methanol | Upto 200 mL |
| Adjust vol to 1L with D/w. | |
| Towbin buffer with SDS, 1L, 10x 25mM Tris, 192 mM glycine, 20% (v/v) methonol (pH 8.3) | |
| Tris base | 3.03 g |
| Glycine | 14.4 g |
| D/w | 500 mL |
| Methanol | Upto 200 mL |
| SDS (10%) soln. | 2.5-10 mL |
Additional Notes
| Methanol - Methanol in the buffer gives efficient binding of proteins to
the membrane. However, methanol also reduces the solubility of proteins in the buffer and also causes
shrinking of the acrylamide gel. Both these negatively affect the elution of the proteins from the gel. The researcher may have to determine (often empirically) the best methanol concentration for the protein and running system that is being used. Generally, for native proteins, methanol is not used. Depending on the sample and membrane, less or no methanol should be used. Recommended percentages of Methanol in buffer are Nitrocellulose - ≤20 % (w/v) PVDF - ≤15 % (w/v) Charged nylon membranes - no methanol |
SDS - SDS is known to improve the elution of the proteins from an SDS gel. Commonly 0.025-0.25% SDS is used. However, SDS reduces the binding of proteins to the membrane. Therefore addition of SDS must be determined for each sample and membrane type individually. |
Native Proteins - For Native proteins it is generally recommended to avoid adding either methanol or SDS. If the proteins are not eluting from teh gel then some amount of SDS might help in the elution but one has to empirically ascertain the functionality of the protein after transfer. |
