Hi Everyone
Im trying to extract and purify bacteriocin from a new species of lactic acid bacteria.
Here, I extracted the bacteriocin from the MRS media using Amberlite XAD-16 (Sigma) and eluted them using 70% isopropanol (200 mL) and later concentrate them using rotary evaporator at low temperature (35 degree celsius) until the volume reach around 25 mL.
However, before proceed to Cation exchage chromatography, I would like to see the protein present. I did the tricine sds page with 4% and 16.5% resolving gel.
I dissolve my sample in sample buffer in a ratio 1:1 and heated at 95 degree celsius for 5 min (constant volt AT120 degree celsius, I followed the recipe from Haider et al., 2012: Haider, S. R., Reid, H. J., & Sharp, B. L. (2012). Tricine-sds-page. In Protein electrophoresis (pp. 81-91). Humana Press, Totowa, NJ.).
The ladder appear good. However, I did not see any band present for my sample ..Can someone suggest/advice what happen here?
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