Description
Endo-off Solution I
Size 10X, 30 mL
Protocol
1. Sample Preparation:
Endotoxin removal is more efficient when the protein concentration is under 2 mg/mL.
Sample buffer pH is better to be between 7 to 8. If you protein pI is around that
range, try pH 4.
Ionic strength of the protein solution is also important. Adjust the salt
concentration to between 300 mM to 500 mM if your protein can tolerate high
salt.
2. Endotoxin Removal Procedures:
1). Take aliquots for analysis (0.5mL)
2). Add Endo-Off Solution to 1 x final
3). Mix by inverting the tube and shaking by hand until
homogenous (~1min)
4). Incubate at 4ºC on a nutator for a minimum of of 30 min
5). Incubate at 37ºC in a waterbath untill solution turns cloudy (3-5 min)
6). Centrifuge in 50mL conical tubes @ 30ºC 10 min 3,313g
7). Aspirate and save the upper aqueous phase
8). Transfer the bottom phase + lower part of the aqueous phase into a 15 mL
conical tube
9). Centrifuge @ 30ºC 10 min 3,313g
10). Aspirate the upper aqueous phase, pool with the aqueous phase collected
in step 7
11) Repeat starting with step 2 if the endotoxin level is still
above the limit.
Note:
1. For small volume sample, like volume < 1 mL, 1.5 mL clean tube and
microcentrifuge are recommended. Just use same centrifugal force and
proceed at same temperature.
2. Wear gloves, protective glasses and lab jacket at all times.
3. All the transfer pipette, tips, tubes and solutions used for endo-removal
have to be sterile and endo-free.
Data Sheet
| Size | 10X, 30 mL |
