Proteomics MS Facility
Handover Date:
Project No.:
Contact Details
Name
CECAD
Group Leader
SFB 635
neither
Institute/Department
Phone
Email
Communication
language
English
German
Sample Details
Organism (for
Main cell / organism:
database search)
Human Derived Samples: no
yes
Additional expressed protein organism:
Expected Proteins (provide amino acid sequences digital as *.txt or *.fasta-file!)
Chemicals used for reduction/alkylation
reduced with:
alkylated with:
not reduced/alkylated
Type of sample
Volume:
pH:
Buffer:
Solution
Gel
Digested
no yes
Used facility protocol
FASP in solution
other:
Used own protocol
in solution
other:
On StageTip:
yes
no
In AS vial:
yes
no
Fractionated by:
OFFGEL
SAX
SCX
other:
Phosphoenrichment:
Gel type
1D
2D;
SDS
native
Excised bands/spots
Gel staining
Coomassie
Silver
other:
Cut into cubes (ca. 1x1 mm)
Amount of protein on gel per lane:
Digested and on StageTip
Gel picture provided?
file
printout
(Please label picture with: Name, Date, MW of Marker bands, Band-/Spot-No.)
Other
Sample Label + Description
1.
2.
3.
4.
5.
6.
7.
8.
Version 1.4
Conc.
Vol.
Facility: sample name / dilution / inj.volume
in µg/µl
in µl
(filled out by facility)
9.
10.
11.
12.
Sample Processing
in gel digestion
in solution digestion
Phosphoenrichment: Fe-NTA TiO2
FASP 3 kDa
FASP 10 kDa
other:
Type of Analysis
LC-ESI-MS/MS: 1 h-Gradient
LC-ESI-MS/MS: 2.5 h-Gradient
LC-ESI-MS/MS: 4 h-Gradient
MALDI-MS: PMF, TOF/TOF
other:
ID
PTMs:
Quantification:
Molecular Mass:
other:
Estimated Costs:
Costs accepted: yes
Collaboration discussed:
yes
no
Mass Spectrometer
Q Exactive Plus
LTQ Orbitrap Discovery
TripleTOF 5600+
4800 Plus MALDI TOF/TOF Analyzer
Remarks
I declare that all samples are free from harmful pathogens, are non-radioactive in nature, and pose no
increased harmful risks to human health.
Samples accepted by
Signature
(facility staff)
(customer)
Version 1.4
Proteomics MS Facility
Sample processing by facility
Samples from:
Handover date:
Chemicals used for reduction/alkylation:
reduced with:
not reduced/alkylated
Enzyme used for digestion:
Lys-C
Digestion protocol:
Trypsin
in gel digestion
other:
alkylated with:
other:
FASP 3 kDa
FASP 10 kDa
FASP 30 kDa
For FASP:
Protein amount (µg) used
for digestion:
Facility sample name
μg enzyme used
1.
2.
3.
4.
Fractionated by:
OFFGEL
Phosphoenrichment
SAX
Fe-NTA
SCX
other:
TiO2
Stage tipping: Each sample loaded on
1,
1-layer (<10 μg)
2-layer (35-70 μg)
2,
more
other:
Stage Tips:
Remarks:
Done by:
Version 1.4
Date:
Proteomics MS Facility
Hardware for measurement
Instruments used for measurement:
Proxeon Easy nLC 1000 coupled to Q Exactive Plus ( QE1
QE2) with Proxeon nano ESISource
Proxeon Easy nLC II coupled to LTQ Orbitrap Discovery with Proxeon nano ESI-Source
Eluents:
A: 0.1 % Formic Acid in H2O
B: 0.1 % Formic Acid in Acetonitrile
Column:
Thermo, Acclaim PepMap RSLC, 50µm x 15cm, nanoViper, C18, 2µm, 100 Å (Q Exactive Plus)
Dr. Maisch, 150 mm x 75 µm, Reprosil-Pur C18-AQ, 3 µm, 120 Å (LTQ Orbitrap Discovery)
Emitters:
Stainless Steel Emitters, length 40 mm, O.D. 150 μm, I.D. 30 μm (ES542)
Sequence:
Remarks:
Version 1.4