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Proteomics and Biochemistry



1.    EQUIPMENT
2.    PROTEIN EXPRESSION AND PURIFICATION
3.    SERVICES OFFERED
4.    RATES
5.    FACILITY CONTACT
6.    UPO LIBRARY RESOURCES
7.    EDUCATIONAL WEB PAGES



 

1. EQUIPMENT

The Proteomics and Biochemistry Unit is located in the Andalusian Centre for Developmental Biology and is equipped with the following instruments:

      
a)    Miniprotean® Tetra-cell (Bio-Rad)
 
Electrophoresis cell for 1-D or 2-D gel electrophoresis to run up to four
minigels (7 x 8 cm) at the same time.
   

b)    IPGphor (GE Healthcare)
 
 
Unit for Isoelectric focusing equipped with individual Strip Holders (6 of 24 cm and 6 of  7 cm long) and a Manifold Unit (to run up to 12  Immobiline DryStrips from 7 to 24 cm at the same time).


c) Ettan Dalt six (GE Healthcare)

Electrophoresis cell for 2-D able to run up to
6 maxi gels (24 x 20 cm) at the same time.


d) ÄKTA Explorer (GE Healthcare)

Platform for protein purification


e) BiacoreTM X100 (GE Healthcare)



f) Typhoon 9410 and Image scanner III (GE Healthcare)  

 

g) ChemiDoc XRS
 

h) Image acquisition and data analysis softwares
 
DeCyder v 7.0
ImageQuant v 5.2
ImageMaster 2D Platinum v 6.0 and 7.0
ImageScanner LabScan 6.0
ImageLab 5.0
 


2. PROTEIN EXPRESSION AND PURIFICATION:

The facility offers a collection of vectors and bacterial strains for the expression and purification of recombinant proteins to the CABD community. We also give technical assistance on the complete purification process.

PLASMID COLLECTION

Name Tag Location
pET30a+ 6His 02
pET9a   03
pET9b   04
pET9c   05
pET9d   06
pGEX 3X GST 10
pGEX 2T GST 11
pGEX 5X1 GST 12
pTXB1 Intein 13
pTYB21 Intein 14
pET14b 6His 15
pQE30 6His 16


BACTERIAL STRAINS COLLECTION

Name Genotype Location
BL21 DE3 pLysS F– ompT hsdS(rB– mB–) gal dcm λ(DE3) pLysS (CamR )(λ(DE3): lacI, lacUV5-T7 gene 1, ind1, sam7, nin5 ) 01
NCM631-pIZ227 hsdS gal _DE3:lacI, lacUV5::gen1 (T7 RNA polymerase) _lac linked to Tn10. 07
Rossetta pRare F– ompT hsdSB(rB– mB–) gal dcm (DE3) pRARE2 (CamR) 08
Rossetta pRare pLysS F– ompT hsdSB(rB– mB–) gal dcm (DE3) pLysSRARE2 (CamR) 09


MANUALS covering several aspects of protein expression and purification techniques:

-GST handbook
-pET system manual
-QIAexpressionist
-GE Recombinant Protein Purification Handbook
-GE Strategies for Protein Purification
-Impact System for intein fusions
-Rosetta cells


2. SERVICES OFFERED:

- Technical assistance for your Proteomic and/or Biochemical experiment which can include sample preparation, protein gel electrophoresis, protein purification or protein-protein interaction analysis.

- Training in the use of instruments or acquisition images softwares for students or researchers.
 
- Training in protein electrophoresis, enzymatic assays, protein purification and protein interaction analysis.

- Antibody purification from serum.


3. RATES:

 

Rates (€, IVA not included)

 

 

 

 

CABD-UPO

OPIs

Private

Sample preparation

 

 

 

Chemical lysis

1

1.5

2

Chemical lysis plus grinding

7.5

9.5

11.5

Sample cleaning (1)

5.5

7

9

Standard quantitation (2)

1.5

2

2.5

Protein quantitation (3)

1

1.5

2

 

 

 

 

1-D electrophoresis

 

 

 

Minigel (8x7 cm)

1

1.5

2

Tank Buffer 1L

2.5

3.5

4

 

 

 

 

2-D electrophoresis (4)

 

 

 

Minigel (8x7 cm)

10

12.5

15.5

Midigel (10-18 x 22 cm)

17.5

22

27.5

Maxigel (26x22 cm)

25

31.5

39.5

DIGE maxigel(6)

270

337.5

422

IEF with Manifold (1-12 strips)

8

10

12.5

Tank Buffer 1L

2.5

3.5

4

Gel staining (4)

 

 

 

Coomassie minigel

3.5

4.5

5.5

Coomassie maxigel

15

19

22.5

Fluorescent staining minigel

16

20

25

Fluorescent staining maxigel

158

198

247

Silver staining minigel

4

5

6.5

Silver staining midigel

15

19

23.5

Silver staining maxigel

18.5

23.5

29

 

 

 

 

FPLC (5)

 

 

 

Day of use

5

6.5

8

 

 

 

 

BIACORE (5)

 

 

 

Day of use

10

12.5

16

 

 

 

 

 

 

 

 

(1) Cleaning is made with the 2-D Clean Up kit. Solubilization buffers are included in the price

(2) Standard quantitation is needed once for each quantitation

 

(3) Price is for one sample with two replicas.

 

 

(4) Prices include Loading buffers

 

 

 

(5) Chemicals, columns, chips, etc…are not included in the price

 

(6) Image acquisition and data analysis included

 

 

 


4. FACILITY CONTACT 

Contact with Laura Tomás
Telephone: 34-95 4978208
E-mail: ltomgal(at)upo(dot)es


5. UPO LIBRARY RESOURCES

a) Books

  • Separation methods in proteomics. Smejkal, Gary B., Lazarev, Alexander. ISBN 0-8247-2699-5. Boca Raton: CRC Press, 2005. (547 SEP)
  • New and emerging proteomic techniques. Dobrin Nedelkov and Randall W. Nelson. ISBN 1-58829-519-2. Totowa (New Jersey, USA): Humana Press, 2006. (575 NEW)
  • 2-D proteome analysis protocols. Andrew J. L. ISBN 0-89603-524-7. Totowa (New Jersey): Humana Press, cop. 1999. (577 PRO)
  • Posttranslational modifications of proteins: tools for functional proteomics. Kannicht, C. ISBN 0-89603-678-2. Totowa (New Jersey): Humana Press, cop. 2002. (577 POS)
  • Proteomics in practice: a laboratory manual of proteome analysis. Westermeier, R. ISBN 3-527-30354-5. Weinheim: Wiley, cop. 2002. (577 WES pro)
  • Proteomics. O'Connor, C.D. & Hames, B.D. ISBN 978-1-904842-13-2. Scion Publishing. 2006. (577 PRO)


b) e-Books

  • Proteomics from protein sequence to function.  Pennington, S.R. and Dunn, S.J. ISBN 1859962963. Oxford: BIOS, 2001.
  • Proteome analysis: interpreting the genome. David Speicher, The Wistar Institute, Philadelphia, PA 19104. ISBN: 978-0-444-51024-2. Elsevier, 2004.


6. EDUCATIONAL WEB PAGES

- Expasy
- Uniprot
- Biacore System
- 2-D electrophoresis and DIGE. GE Healthcare
- All about Biacore consumable catalogue.