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TEL: +32 16 58 90 45
Fax :+ 32 16 50 90 45
GENTAUR Europe
tel+32 2 732 5688
fax+32 2 732 4414
Av.
de l' Armée 68
B-1040 BRUSSELS
BELGIUM

GENTAUR France
tel 01 43 25
01 50
fax01 43 25
01 60
9, rue
Lagrange
75005 PARIS
FRANCE

GENTAUR Italy
tel 02 36 00
65 93
fax 02 36 00
65 94
20135 MILANO
ITALY

GENTAUR Germany
tel +49 241
6085 13140
fax +49 241
6085 33033
Forckenbeckstraße 6,
D-52074
Aachen
GERMANY
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Storage Logistics
You provide us
with a,b, or c:
a)
Homogenous protein in an excised band from a dried SDS-gel (stained
with Coomassie Blue or Silver, preferably without fixation). Please
enclose a gel photo. Please mention the %age of the gel, clearly mark
the band which was excised from the gel, and specify the molecular
weight marker and the estimated molecular weight of the protein of
interest.
b)
Homogenous protein in an excised spot from a dried 2-D-Gel (stained
with Coomassie Blue or Silver, preferably without fixation). Please
enclose a gel photo. Please clearly mark the spot which was excised
from the gel.
c) At
least 5 µg purified native protein (ammonium sulfate precipitate or
lyophilizd protein). Please enclose a gel photo (SDS-PAGE), mention
the %age of the gel, clearly mark the band which corresponds to the
purified protein, and specify the molecular weight marker and the
estimated molecular weight of the protein of interest. Recommend a
suitable buffer for dialyzing (ammonium sulfate precipitate) or
dissolving (lyophilized material) the protein. If this is not a
standard buffer, add your buffer in sufficient amount (esp. for
dialysis). Mention special requirements of your protein with respect
to solubility and stability (especially important for membrane
proteins).
Required
Information:
Source of the
protein (organism, tissue, cell fraction etc.) and, if possible, any
assumption about the nature and/or function of the protein of
interest.
If available: Molecular Weight of protein and its subunits, and
isoelectrical point.
Please ship the
protein without cooling (dried gel bands or spots, lyophilized protein)
or on blue ice (ammonium sulfate precipitate) to:
Gentaur bvba
Avenue de l'Armée 68/B4,
B-1040 Brussels
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Our service:
- (In gel)
trypsination of the protein (band or spot) of interest (protein is
cleaved at arg and lys sites into peptides)
- MALDI-TOF Mass
Spectometry analysis of the tryptic peptides
- Protein
identification by computational tools (database searching combined
with Bayesian statistics): Masses of peptides from the proteolytic
digestion of the corresponding protein are compared to the masses of a
peptide database based on the non redundant information extracted from
NCBI, GenBank, CDS translations, PDB, SwissProt, PIR, and PRF protein
databases. The result is a ranking of candidate proteins based on
their calculated posterior probability. The protein identity is
re-verified using pI, MW and peptide mass fingerprinting data and
comparing them with the theoretical peptides calculated for all
proteins of question in the SWISS-PROT/TrEMBL databases. This
verification process is carried out by trained scientists who verify
the protein identity by checking the peptide pattern peak by peak for
plausibility taking trypsin autolysis and trypsin cleavage faults into
account.
- You receive a
detailed analysis report featuring the identity of the protein and the
likelihood of the correctness of the identification. In case it is a
new protein, the features of similar/related proteins are listed.
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Order
Information:
Cat. Nr.
PIPB01: Protein Identification by MALDI-TOF-MS, protein shipped in
protein gel band
Price: 500 Euro
Cat. Nr.
PIPS01: Protein Identification by MALDI-TOF-MS, protein shipped in
2 D gel spot
Price: 500 Euro
Cat. Nr.
PIPP01: Protein Identification by MALDI-TOF-MS, purified protein
provided
Price: 570 Euro
Please note:
In case the protein amount or purity was not sufficient to perform
MALDI-TOF MS, a set up fee of € 175 is charged. |
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| Examples: |
| The
SDS-PAGE gel illustration below shows the results of a protein
purification (upper band) by metal-chelate chromatography after trying
different washing conditions applied to eliminate the lower band
(impurity). |
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| The
MALDI-TOF MS spectrum of the protein preparation confirmed the results
obtained with the SDS-PAGE. |
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| The contaminant
has a molecular weight of 37095 Da. To efficiently eliminate the
impurity, the 37 kDa protein was identified by MALDI-TOF MS of a tryptic
protein digest:
The lower band was
excised from the gel and the in gel-digestion was carried out with
trypsin. The tryptic digest was analysed with MALDI-TOF MS. The
resulting mass spectra are given below: |
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Protein Candidates |
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Rank
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Probability
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Protein
Description |
MW (kDa)
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1
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1.0e+00
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gi|6324486|ref|NP_014555.1| Alcohol dehydrogenase; Adh1p [Saccharomyces
cerevisiae] |
37
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2
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3.2e-03
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gi|1168350|sp|P00330|ADH1_YEAST ALCOHOL DEHYDROGENASE I |
37
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3
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7.8e-09
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gi|6324174|ref|NP_014244.1| Ynl155wp [Saccharomyces cerevisiae] |
32
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4
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1.1e-09
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gi|7245674|pdb|1YLV|A
Chain A, Schiff-Base Complex Of Yeast 5-Aminolaevulinic Acid Dehydratase
With Laevulinic Acid |
38
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5
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3.9e-10
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gi|171974|gb|AAA34790.1| (M81696) mitochondrial ribosomal protein
[Saccharomyces cerevisiae] |
29
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6
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3.2e-10
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gi|6325350|ref|NP_015418.1| Ypr093cp [Saccharomyces cerevisiae] |
36
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7
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3.0e-10
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gi|223142|prf||0512226A dehydrogenase isozyme I,alcohol [Saccharomyces
cerevisiae] |
32
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8
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3.0e-10
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gi|6320613|ref|NP_010693.1| 263-amino acid mitochondrial ribosomal large
subunit protein; similar to L23 family of ribosomal proteins; Mrp20p
[Saccharomyces cerevisiae] |
31
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9
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1.2e-10
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gi|626218|pir||S45958
probable membrane protein YBR090c - yeast (Saccharomyces cerevisiae) |
13
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10
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1.2e-10
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gi|6319560|ref|NP_009642.1| mitochondrial ADP/ATP translocator; Aac3p
[Saccharomyces cerevisiae] |
33
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The contaminating
protein was identified as alcohol dehydrogenase and could now be removed
easily from the mixture by exploiting its known hydrophobicity (washing
the column with isopropanol). |
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Our service:
Sample
preparation and MALDI-TOF Mass Spectrometry of the protein.
Determination of the absolute mass of the protein subunit(s).
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Order
Information:
Cat. Nr.
DMW01
Absolute Molecular Weight Determination of purified protein by
MALDI-TOF MS
Price: 225 Euro
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