Description
Protein extraction from cell wall-containing microbes is a frequent procedure in bio-research
Labs. The methods for protein extraction from these microbes are usually harsh, tedious and not
sufficiently reliable. YT-015 provides an instrument-free, rapid and gentle way for extracting
proteins from microbes with thick and strong cell walls. These microbes include but not limited
to yeast, filamentous fungus, gram positive and negative bacteria, insect eggs and microalgae.
This kit contains optimized denaturing and native protein extraction buffers for user to choose
from. Unlike many other methods of which harsh conditions, such as 8 M urea, glass bead lysis
using an instrument, and boiling the sample with alkaline extraction etc., are used for yeast
protein extraction. Proteins in certain molecular weight range are usually preferentially extracted
with these methods. This kit features a single tube protocol, full range of yeast proteins are
extracted without bias. The kit works well for both log phase and stationary phase microbes. The
whole procedure takes less than 10 min to complete and the protein yield is in the range of 2-5
mg/ml. The materials provided are sufficient for 50 extractions.
Application
Proteins extracted with this kit can be directly used for many downstream applications such as
SDS-PAGE analysis, Western blotting, IP. ELISA and enzyme activity assays and proteomic
analysis. The buffers are compatible with IMAC resins for his-tagged protein purification
Kit components
1. 20 ml Denaturing Buffer
2. 20 ml Native Buffer
3. 5 g protein extraction powder
4. 4 pestles for 1.5 ml microcentrifuge tube
5. 1.5 ml microfuge tube X 50
Storage: Store the kit at room temperature.
Additional Materials Required
Table-Top Microcentrifuge with a maximum rpm of 14,000-16,000.
Important Product Information The use of protease inhibitors is not necessary prior to extraction. However if downstream application takes significant amounts of time or the protein extract will be stored for longer period of time, addition of protease inhibitors to extraction buffer is recommended. For determination of protein concentration, BCA kit (Pierce) isrecommended. To study protein phosphorylation, phosphatase inhibitors (such as PhosStop from Roche) should be added to the buffer prior to use