Protein extraction from fungi and bacteria can be challenging, due to the stable cell wall which must be disrupted prior to detergent-assisted protein extraction and solubilization.

The TOTAL protein extraction kit has been specially formulated for efficient extraction and solubilization of total protein mixtures from fungi and bacteria.

After mechanical disruption of the cell wall by glass beads, proteins are solubilized using a complex detergent mix. The use of several detergents ensures uniform solubilization of proteins with widely different characteristics, including strongly hydrophobic proteins, such as integral membrane proteins.

After solubilization, proteins are denatured by addition of sample buffer and loaded onto an SDS-PAGE gel for further analysis.

The kit is recommended for preparation of total protein extracts in DUALhybrid, DUALmembrane and DUALhunter screens and is also compatible with  other yeast two-hybrid systems.


Comparison of extraction efficiencies using the TOTAL protein extraction kit. 30 mL stationary phase yeast cultures expressing a DUALmembrane  EGFR bait were extracted using either the TOTAL protein extraction kit or two different protein extraction kits from competitors. Lane 1: Molecular weight marker (10-250 kDa), Lanes 2 and 3: two independent extractions using the TOTAL protein extraction kit, Lane 3: kit from competitor C, Lane 4: kit from competitor P.

 

Kit contents

Extraction buffer H (20 mL)      
Sample buffer (3 mL)     
Glass beads (12 mL)     

Sufficient for 60 extractions of 30 mL cultures

 

 

Related products

P01001   DUALmembrane kit 3

P01004   DUALhybrid kit

P01005   DUALhunter kit

P01006   KickStart protein expression kit

 

 

Order number	Product	Quantity
P01013	TOTAL protein extraction kit	1 kit