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Idiomas disponibles
English
Intended Use
The Auto2D
Electrophoresis Device fully
®
automates the process of 2D electrophoresis,
separating proteins first by isoelectric point and
second by molecular weight. Complete protein
separation is achieved in just two hours.
Contents
• Auto2D
Electrophoresis Device
®
• Electrical Power Cables:
• Power cable for JP
and North America
• 3P-2P Conversion Plug
for JP and North America
• Power cable for Europe
• Power cable for China
• Power cable for UK
Also Needed
(not included)
See Product Ordering on p. 51 for
catalogue numbers.
• Auto2D
Electrode Chip
®
• Auto2D
PAGE Chip
®
• Auto2D
IEF Chip
®
• Auto2D
Solution Chip
®
• Auto2D
Tris-Glycine or Tris-Tricine
®
Reagent Kit
• Ampholyte (choose appropriate
ampholyte based on IEF Chip pH range)
• Distilled water
User Guide Online
The complete User Guide includes detailed
instructions and recipes. It can be downloaded
from the Auto2D
Device product page at
®
SigmaAldrich.com.
Storage and Stability
The Auto2D
device should be used/stored
®
indoors only. See each component's product
label for storage requirements.
20424817 Rev 06/21
All manuals and user guides at all-guides.com
Precautions
Y
Y
For research use only.
Please read the Safety Sheet (enclosed) and
the complete User Guide at
SigmaAldrich.com
carefully before using this product.
Auto2D
Workflow
®
Total process: 100–155 minutes
(depending on the recipe)
Sample Preparation
Removal of Impurities
Addition of Reagents
Sample Loading
30 Minutes
and Gel Rehydration
5 Minutes
First Dimension:
30 Minutes
Isoelectric focusing
SDS Equilibration
5 Minutes
Second Dimension:
30 Minutes
SDS-PAGE
Detection
2
Preparation
Reagents
1.
Prepare and store Reagent Kits as
directed in the literature provided with the
kits, or the complete User Guide (online).
2.
Allow the following products stored at
low temperatures to equilibrate to room
temperature (20–25 °C approximately
10 minutes).
• IEF Chip
• PAGE Chip
• Rehydration Solution
• DTT Solution
• Ampholyte
3.
Prepare Working Rehydration Solution
for rehydration of IEF Chip and
extraction/dilution of protein sample:
Reagents
Rehydration
Solution
DTT Solution
(1M)
Ampholyte*
Total
* Select ampholyte according to the range
of IEF Chip used. For ampholytes at
40% w/v stock concentration, add 1 µL.
For ampholytes at 100X, add 2 µL.
4.
Prepare Working Equilibration Buffer for
equilibration of focused proteins before
SDS-PAGE.
Reagents
Equilibration
buffer premix
DTT Solution
(1M)
Total
English
20424817 Rev 06/21
Sample Preparation
Dissolve the protein sample in the
Working Rehydration Solution, prepared
according to step 3. Sample may be
diluted 2-fold or more with Working
Rehydration Solution to reach desired protein
concentration and decrease salt concentration.
A high salt concentration can affect protein
separation during isoelectric focusing and
cause the current to exceed 100 µA. Samples
with a high salt concentration should be
desalted by either:
TCA/acetone precipitation and resuspension
of proteins in Working Rehydration Solution
OR
Buffer exchange using a spin column
NOTE: During protein resuspension, avoid
heating proteins over 37 °C.
Quantify proteins. Between 0.1–25 µg of
Final
protein can be loaded for 2D electrophoresis.
Volume
Concentration
The optimum amount of protein will depend on
detection method and sample complexity. The
189 µL
following protein amounts should be used as a
starting point, users may need to optimize the
10 µL
50 mM
loading amount for their particular sample.
• Coomassie Brilliant Blue staining: 25 µg
1–2 µL
0.5–1% v/v
• Fluorescent staining: 10 µg
200 µL
• Silver staining: 5 µg
• Fluorescent pre-labeling: 3 µg
The sample volume applied to the device
is 10–12 µL. Sample can be diluted with
Working Rehydration Solution as necessary.
Power the Auto2D
Final
Select the appropriate power cord (provided
Volume
Concentration
with the device) that is compatible with the
power outlets in your country. Firmly push
760 µL
the matching end into the back of the
Auto2D
Device, and the other end into
®
40 µL
50 mM
the power outlet.
800 µL
The power switch is located next to the AC plug
port on the back of the Auto2D
the switch up to turn on.
The application will be automatically launched.
From the screen, select Auto2D
(original).
3
Device
®
Device. Press
®
®
mode
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