Download KUMASI™ stabilized colloidal stain for polyacrylamide gels

KUMASI™
KUMASI™ stabilized
colloidal stain for
polyacrylamide gels
The Coomassie Chronicles
The Coomassie dyes, named in commemoration of
the British occupation of the Ashanti capital Kumasi in
Ghana, have remained in use for over 50 years. Faseka
de St. Groth et al [1] first described the use of Coomassie
Brilliant Blue (CBB) for staining proteins separated on
cellulose acetate membranes and demonstrated its strict
adherence to Beer’s law. Soon afterwards, CBB would
be popularized for staining proteins in polyacrylamide
gels [2,3]. Members of the first Electrophoresis Societies
became known as the “Bluefingers” and some German
practitioners were mistaken for counterfeitors for the
residual blue stain always on their hands.
Comparison of CBB to silver staining
Silver stain is inarguably the most sensitive stain. It is
mimics a photographic process in which the latent image
is “developed” by the reduction of silver ions. However,
since the binding of silver ions to proteins is highly
dependent on amino acid composition, all proteins are
not stained equally and some are poorly stained or not
stained at all. There is a low correlation between silver
staining and CBB, where r = 0.657.
KUMASI stabilized colloidal Coomassie
stained two-dimensional gels with greater
sensitivity than the SYPRO Ruby and
Deep Purple fluorescent stains.
KUMASI stabilized colloidal stain can be
reused at least four times, reducing waste
streams by 75% or more.
Comparison of KUMASI to fluorescent dyes
KUMASI stabilized colloidal Coomassie stained twodimensional gels with greater sensitivity than the SYPRO
Ruby and Deep Purple fluorescent stains (Figure 1).
Further, the photoinstability of these stains has been
reported [6]. Deep Purple fluorescence decreases nearly
30% after only two minutes of UV transillumination.
The correlation between SYPRO Ruby and KUMASI
is r = 0.825.
Neuhoff et al. [4] and Candiano et al. [5] exploited the
colloidal properties of the dimethylated form of CBB and
achieved sensitivity similar to silver staining.
Figure 1. Relative sensitivity of KUMASI (left), SYPRO Ruby
(center) and Deep Purple stains. Mean number of protein
spots detected in triplicate gels is reported (lower left). Insets
show differential staining of proteins.
References
1. Fazekas De St. Groth S, Webster RG, Datyner A. (1963). Biochim. Biophys. Acta. 71, 377-385.
2. Meyer TS, Lambert BL (1965). Biochim. Biophys. Acta. 107, 144-145.
3. Altschul, A. M.; Evans, W. J. (1967). Methods in Enzymology 11, 179–186.
4. Neuhoff V, Stamm R, Eibl H. (1985). Electrophoresis, 6, 427-448.
5. Candiano G, Bruschi M, Musante L, Santucci L, Ghiggeri GM, Carnemolla B, Orecchia P, Zardi L, Righetti PG.
Electrophoresis 25, 1327-1333.
6. Smejkal GB, Robinson MH, Lazarev A. (2004). Electrophoresis 25, 2511-2519.
Comparison to competing Coomassie Dyes
KUMASI stabilized colloidal stain detected 28% more protein spots in
two-dimensional gels of Escherichia coli lysate than Thermo-Pierce GelCode
Blue Stain (Figure 2).
Reducing your Coomassie footprint
Compared to conventional Coomassie stains which generate chemically hazardous
waste streams, the new generation of “ecologically friendly” Coomassie stains
exhibit significantly decreased sensitivity. The solution is the KUMASI stabilized
colloidal stain which can be reused at least four times, reducing waste streams by
75% or more. The KUMASI Fixative and Enhancer solutions can also be reused
and are compatible with sink disposable.
Figure 2. Identical 2D gels of Escherichia coli lysate stained with
KUMASI (left) or GelCode Blue (right). The mean number of total protein
spots detected in duplicate gels is reported (upper right). KUMASI
detected 219 proteins not detected by GelCode Blue.
Focus Proteomics is a proteomics
CRO specializing in protein fractionation,
two-dimensional gel electrophoresis
(2DGE), and label-free quantitative
mass spectrometry (LFQMS) to identify
and quantify proteins and their posttranslational modifications.
Contact: Phone:
Email:
Focus Proteomics, LLC
32 Overlook Circle
Hudson, NH 03051
(603) 204-4947
[email protected]
www.focusproteomics.com