My workflow would be as follows, as I tested it and found the most satisfying for creating signatures for emails:
- Preferably use a graphic tablet and a stylus to create a hand signature
- In Illustrator create a regular A4/Letter document
- Use the Brush tool: Caligraphic Brush, e.g. 5pt Oval
- Draw a large, comfortable signature
- Turn on Window>Pixel View and adjust the size of the signature for your final output. After scaling down the path, the 5pt brush might be too large, you can either pick smaller brush 3pt or manipulate with the path/stroke size. Keep it a bit thicker rather than thinner, it will look nicer.
- Save for web: GIF, Grayscale, Transparency
Voila. Good luck.