ProteoSpin™
Detergent Clean-up Kits
Detergents are
extensively used to prepare protein samples. However, detergents
must often be removed prior to downstream analysis because of their
undesirable effects. These include extraneous peaks in mass
spectrometry, artifacts with chromatography and electrophoresis,
interference with microinjection into cells and interference with
protein immunization.
The ProteoSpin™
Detergent Clean-up Micro Kit provides a fast and simple procedure
for the effective removal of SDS, Triton X-100 and other detergents
from small samples up to 50 mg of protein. The kit employs an
innovative separation material, silicon carbide (SiC). Processed
using proprietary technology, SiC acts as an ion exchanger.
ProteoSpin™
Detergent Clean-up Kit Benefits
-
A complete kit for acidic and
basic protein samples
- The ProteoSpin™ Detergent Clean-up Micro Kit contains necessary
solutions for processing small detergent-containing samples from
2–50 mg of protein. Solutions for processing both acidic and basic
proteins are included.
-
No detergent carryover
- Unlike HPLC procedures, each sample is processed individually
using its own spin tube providing no carryover or column bleed
from sample to sample.
-
Useable with a variety of
detergents -
The kit can be used to remove detergents including SDS, Triton
X-100, CHAPS, NP-40, and Tween 20.
-
Effective detergent removal
- Removes more than 95% of detergents for many protein samples,
allowing tryptic digestion.
-
High recovery
- Protein recoveries of 80 to 95% for many proteins.
Fast processing
time
- In contrast to longer dialysis procedures, the easy-to-use
procedure allows processing up to 12 samples in only 20 minutes.
Each ProteoSpin™
Detergent Clean-up Micro Kit spin tube contains 25 mg SiC. Capacity
per spin tube is approximately 2-50µg and may vary depending on
protein sample.
ProteoSpin™
Detergent Clean-up Micro Kit Contents
Each ProteoSpin™
Detergent Clean-up Micro Kit provides essential components for use
with the provided protocols.
-
Column activation and wash
buffer for acidic proteins
-
pH binding buffer for acidic
proteins
-
Column activation and wash
buffer for basic proteins
-
pH binding buffer for basic
proteins
-
Elution buffer
-
Neutralizer
-
ProteoSpin™ SiC micro spin
columns
-
Collection tubes (2 mL
capacity)
-
Final collection tubes (1.7 mL
capacity)
-
ProteoSpin™ Detergent Clean-up
Micro Kit application manual
-
Protocol reference card for
both acidic and basic procedures
Customer-supplied
Reagents and Equipment
-
Benchtop microcentrifuge
-
Micropipettors
-
pH indicator paper
-
Other optional elution buffers
-
Isopropanol
Technical Note
Abstract
The efficacy of the
ProteoSpin™ Detergent Clean-up Kit to
remove the commonly
used non-ionic detergent Triton X-100 from a protein sample was
investigated. For comparison, the efficacy of an ultrafiltration
membrane that removes detergent based on size exclusion was also
investigated.
The ProteoSpin™
procedure took approximately 20 minutes to complete and resulted in
a recovery of 71.6% of the protein. No Triton-X 100 peaks were
observed by mass spectrometry analysis, indicating that the
ProteoSpin™ method effectively removed the Triton-X 100. In
contrast, the ultrafiltration procedure took 2 hours to complete and
the percent recovery could not be determined since residual
detergent in the sample interferes with the BioRad protein assay.
Ultrafiltration-processed protein was found to contain a series of
characteristic Triton X-100 peaks.
Since Triton X-100
is inhibitory to trypsin digestion, purified protein solutions were
mixed with trypsin and the resulting reaction mixtures were analyzed
by SDS-PAGE. SDS-PAGE analysis demonstrated that the majority of
protein recovered using the ProteoSpin™ procedure was digested by
trypsin. In contrast, a substantial amount of protein recovered from
the ultrafiltration membrane remained undigested.
Table 1 illustrates results of an experiment that examined the
removal of the anionic detergent SDS with the ProteoSpin™ Detergent
Clean-up Kit. The procedure removed more than 99% of the SDS from
the protein sample.
Downloadable
Detergent Removal Kit Application Notes
Removal of SDS
Removal of Triton
Downloadable Instruction Manual
Detergent Removal
Manual
