PRODUCT INFORMATION
HyTra Cell Protein Extraction Reagent
Product No. W-7850 / 500ml
Introduction
HyTra Cell Protein Extraction Reagent is ready-to-used reagent for total
protein extraction from cell cultured mammalian cells. The protein extracts
can be used for reporter gene expression assays (β-gal, CAT, alkaline
phosphatase), immunoassays (Western blots, immunoprecipitation, ELISA),
kinase assays (PKC, tyrosine kinase), and phosphatases (general
phosphatases, tyrosine phosphatases). It is compatible with Coomassie Blue
and silver staining. Protein lysates can be used also for DNA-protein
interaction assays (gel-shift assays). Additionally, this reagent can be used
with protease inhibitor (#HC100-007) and/or phosphatase inhibitor
(#HC100-008) if necessary.
Storage
This product is stable for one year at 4℃.
Additional Materials required
‧Protease Inhibitor Cocktail (#HC100-007)
‧Phosphatase Inhibitor Cocktail (#HC100-008)
‧PBS
‧Micropipettes and tips
‧Centrifuge tubes
‧4℃ centrifuge
Protocol

Prepare 4℃ centrifuge.


Keep all samples on ice during operation.
1ml Hytra Cell Protein Extraction reagent is used for 5X107 to 1X108

cells.
If desired, add protease inhibitors and/or phosphatase inhibitor to the
Hytra Cell Protein Extraction Reagent just before use.
Procedure for adhesion Cells
1. Carefully remove (decant) culture medium from adherent cells.
2. Wash cells once in wash buffer (e.g., PBS).
PRODUCT INFORMATION
3. Discard PBS.
4. Add the appropriate amount of Hytra Cell Protein Extraction Reagent to
5.
6.
7.
each plate well. Shake gently for 10 minutes.
Collect the lysate cell by scraping and transfer the cell lysate to a
microcentrifuge tube
Centrifuge the sample at 10,000 × g for 5 minutes to pellet cell debris.
Collect supernatant and continue with downstream analysis or further
purification.
Procedure for suspension cells
1. Collect the cells into an appropriate centrifuge tube.
2. Centrifuge for 5 minutes at 450 × g.
3. Decant and discard the supernatant.
4. Wash the cell pellets once with PBS and centrifuge for 5 minutes at 450
× g for 5 minutes. Decant and discard supernatant.
5. Resuspend the cell pellet with appropriate amount of Hytra cell protein
6.
7.
8.
extraction reagent.
Keep on ice for 10 minutes, and vortex at 2 mininutes intervals.
Centrifuge the sample at 10,000 × g for 5 minutes to pellet cell debris.
Collect supernatant and continue with downstream analysis or further
purification.
Note: Lysate preservation requires low temperatures. For long term storage it
is recommended to store the lysate at –70 °C.
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