|
|
GENTAUR
 +32
1658 9045
or
0032 (0)16 41 44
07
 +32
1650 9045
info@gentaur.com
Av. de
l' Armée 68
B-1040
Brussels
BELGIUM
France
tel 01 43 25 01 50
fax01 43 25 01 60
9, rue Lagrange
75005 Paris
Italia
tel 02 36 00 65 93
fax 02 36 00 65 94
20135 Milano
Deutschland
tel +32
1658 9045
fax +32
1650 9045
Polska
Tel 00 48 51 760 77
08
Fax 00 32 16 50 90 45
Ulica Ogarna 15/19 B
m2
80-826 Gdansk
日本
tel +81 78 386 0860
fax +81 78 306 0296
Minaatojimaminami-manchi
Chuo-ku, Kobe
065-0047
Österreich
+43720880899
Canada Montreal
+15149077481
Česká republika
Praha
+420246019719
Danmark
+4569918806
Finland Helsset
+358942419041
Ελλάς Αθήνα
+302111768494
Magyarország
Budapest
+3619980547
Ireland Dublin
+35316526556
Luxembourg
+35220880274
Nederland
+31208080893
Norge Oslo
+4721031366
Polska Warszawa
+48223988221
Sverige Stockholm
+46852503438
Schweiz Züri
+41435006251
US New York
+17185132983
Other Countries
0032 (0)16 41 44 07
|
|
|
| |
|
|
|
|
The SMARTTM
Bacterial Protein Extraction Solution is
designed for fast and easy extraction of total
proteins from bacteria without the need for
sonication or precipitation. This solution
utilizes a proprietary non-ionic detergent in
20mM Tris/HCl(pH7.5). In addition, the SMARTTM
Bacterial Protein Extraction Solution offers
several folding increase in the yield of soluble
protein when compared with other commercial
lysis reagent. Depending on the particular
application, additional components, such as
lysozyme, protease inhibitors, salts, reducing
agents and chelating agents may be added to the
solution. The solution may be used for both
soluble protein extraction and inclusion body
purification from bacterial cell lysates. In
fact, yields obtained with SMARTTM
Bacterial Protein Extraction Solution greatly
exceeds those obtained using standard sonication
methods!
Bacteria often over-express recombinant proteins
and form inclusion bodies, which are insoluble
aggregates of misfolded protein. Centrifugation
separates inclusion bodies from soluble
proteins; however, Lysozyme is required for
purification of inclusion bodies. Lysozyme
significantly improves inclusion body purity by
digesting the cell debris. |
|
- Non-ionic
detergent in a Tris/HCl (pH7.5) buffer.
Therefore, use Tris buffer for subsequent
protein purification. Eliminates mechanical
disruption and provides a gentle and
efficient method for extracting proteins.
- Whole
cell lysates prepared with the SMARTTM
Bacterial Protein Extraction Solution are
compatible with the SMARTTM BCA Protein
Assay Kit(Cat.No.21071) and coomassie
protein assays.
|
|
|
* SMARTTM
Bactrial Protein Extraction Solution
|
110 ㎖
|
|
* protease
inhibitor is not included
|
|
1)
|
Upon receipt
store product at room temperature.
|
|
|
 Comparison
of Protein Recovery with Competitor |
 |
Fig. 1.
Comparison of efficiency of protein extraction
with competitor’s Kit.
The results of Table 1 was demonstrated
graphically. The SMARTTM Bacterial
Protein Extraction Solution showed improved
recovery of bacterial protein extraction. |
| |
Table. 1.
Comparison of extracted protein concentration
with competitor
The Quantities of extracted proteins were
estimated using by SMARTTM BCA
Protein Assay Kit (Cat.No 21071)
|
 |
| |
|
Inclusion Body Isolation |
| |
 |
Fig. 4.
Isolated inclusion bodies
Each of inclusion body of several expression
hosts was isolated and solubilized (followed by
Protocol 2).
Lane A,
M15 [rGroEL]; lane B, BL21[rTaq
]; lane C, origami [rTaq];
lane D, Rogettagami [rTaq];
lane M, Protein Marker lane 1,
SMARTTM Bacterial Protein Extraction
Solution; lane 2, Competitor’s
Kit |
|
|
|
|
