Parallel functional protein analysis
With E.coli expressed proteins derived
from cDNA libraries from four different human tissues
(lung, colon, T-lymphocytes,
foetal brain), our sister company Source
BioScience offers one of the largest collection of arrayed proteins
for screening experiments.
Protein array handbook
Search and order
For ordering, please contact Sales.
- Target protein identification, using antibodies and sera
- Functional assays, e.g. phosphorylation, ribosylation,
- Identification of DNA/RNA binding proteins
- Identification of protein−protein interactions
Source BioScience offers the following
2 Protein Array types as standard, readily available products:
- hEXselect - Human random cDNA Protein Array, redundancy
reduced, 24.000 clones - hEXselect protein expression
library is derived from human foetal brain and contains 24,000
clones. It comprises full-length as well as shorter cDNA clones in
an E.coli expression vector. All clones have been 5' tag sequenced
and are fully annotated. The size of the low redundancy collection
allows a complete spotting in duplicates onto 1 Protein Array,
which saves processing time and hybridisation probe volume. Note
that the majority of the clones represent partial proteins
including translated 5' UTR sequences, with one third matching to
the human proteome (Büssow et al., 2000: A human cDNA library for
high-throughput protein expression screening. Genomics. 65(1):
1−8). Protein Arrays are delivered with an accompanying annotation
table and respective spotting positions.
- UniPEx - Human in frame cDNA Protein Array Set,
redundancy reduced, 15.300 clones / 7390 distinct human proteins
- The UniPEx protein expression library consists of 2
Arrays representing clones in 2 different vectors. In total, more
than 100,000 sequenced clones from different protein expression
libraries (human foetal brain, T-cells, lung, colon) were analysed
in depth for their coding potential. After in-frame analysis only
clones with a confirmed in-frame ORF were selected and redundancy
with respect to clones per gene was minimised (< 3 fold). In
total, the 15,300 UniPEx clones are representing 7,390 distinct
human proteins. The UniPEx set comes together with an annotation
table showing cloneIDs per filter position, geneID, gene
description in Excel format, with links to public databases.
- Custom-made Protein Arrays can be generated from
subsets of these libraries
Source BioScience offers a variety of
protein expression clone libraries which are also employed to
create Protein Arrays. Source BioScience Protein Arrays consist of
up to 27,648 spots, which are printed in duplicate (total up to
55,296 protein spots) onto 22 cm x 22 cm PVDF membranes. The
expression vector adds a HIS-Tag to each expressed protein. All
proteins on the arrays have been verified for expression by
detecting this HIS-Tag using an anti-HIS antibody. Protein
expression clone collections and the respective individual cDNA
clones are available for in-depth follow-up-experiments such as
Fig.1 Vector Structure
of Source BioScience protein expression clones
The fabrication of Source BioScience
Protein Arrays combines reliable contact printing technology with
in situ protein expression directly at the surface of PVDF
membranes. Protein synthesis is based on an E. coli based
expression system for recombinant proteins. Expressed proteins are
extracted and immobilised under denaturing conditions, directly at
the PVDF membrane surface. Resulting Source BioScience Protein
Arrays mainly consist of almost linear sequence epitopes, ideally
suited for epitope mapping, for auto antibody assays, and for
antibody cross reactivity screenings
Fig.2 Overview about
Source BioScience's protein array production process and subsequent
screening and analysis procedure
Centre for Human Proteomics
Royal College of Surgeons in Ireland
123 St Stephen's Green
Tel.: +353 1 402 8518
Fax: +353 1 402 8514
Dr. Mark T. Bedford
University of Texas
M.D. A. Cancer Center, Dept.of Carcinogenesis
M. Raab, H. Daxecker, R.J. Edwards, A.
Treumann, D. Murphy, N. Moran.
Protein interactions with the platelet integrin alpha(IIb)
Proteomics (2010) May 18. [Epub ahead of print]
E.W. Dervan, H. Chen, S.L. Ho, N.
Brummel, J. Schmid, D. Toomey, M. Haralambova, E. Gould, D.M.
Wallace, J.H. Prehn, C.J. O'Brien, D. Murphy
Protein macroarray profiling of serum autoantibodies in
Invest Ophthalmol Vis Sci (2010) 51(6):2968-75. Epub 2010 Jan
D. Murphy, J. Parker, M. Zhou, F.M.
Fadlelmola, C. Steidl, A. Karsan, R.D. Gascoyne, H. Chen, D.
Constitutively overexpressed 21 kDa protein in Hodgkin
lymphoma and aggressive non-Hodgkin lymphomas identified as
cytochrome B5b (CYB5B).
Mol Cancer. (2010) 9:14.
D.S. Gibson, J. Banha, D. Penque, L.
Costa, T.P. Conrads, D.J. Cahill, J.K. O'Brien and M.E.
Diagnostic and prognostic biomarker discovery strategies
for autoimmune disorders.
Journal of Proteomics (2010) 73, 1045-1060
D.J. O'Connell, M.C. Bauer, J. O'Brien,
W.M. Johnson, C.A. Divizio, S.L. O'Kane, T. Berggård, A. Merino,
K.S. Åkerfeldt, S. Linse and D.J. Cahill
Integrated protein array screening and high throughput
validation of 70 novel neural calmodulin binding
Mol Cell Proteomics (2010) Jan 12. [Epub ahead of print]
G. Kijanka, S. Hector, E.W. Kay, F.
Murray, R. Cummins, D. Murphy, B.D. MacCraith, J.H.M. Prehn, D
Human IgG antibody profiles differentiate between
symptomatic patients with and without colorectal
Gut (2010) 59: 69-78
E. Kowenz-Leutz, O. Pless, G. Dittmar,
M. Knoblich and A. Leutz
Crosstalk between C/EBPß phosphorylation, arginine
methylation, and SWI/SNF/Mediator implies an indexing transcription
The EMBO Journal (2010) 29, 1105-1115
G. Kijanka, S. IpCho, S. Baars, H.
Chen, K. Hadley, A. Beveridge, E. Gould and D. Murphy
Rapid characterisation of binding specificity and
cross-reactivity of antibodies using recombinant human protein
Journal of Immunological Methods (2009) 340, 132-137
G. Kijanka, R. Barry, H. Chen, E.
Gould, S.K. Seidlits, J. Schmid, M. Morgan, D.Y. Mason, J. Cordell
and D. Murphy
Defining the molecular target of an antibody derived from
nuclear extract of Jurkat cells using protein
Analytical Biochemistry (2009) 395 , 119-124
G. Kijanka and D. Murphy
Protein arrays as tools for serum autoantibody marker
discovery in cancer.
Journal of Proteomics (2009) 72, 936 - 944
O. Pless, E. Kowenz-Leutz, M. Knoblich,
J. Lausen, M. Beyermann, M.J. Walsh and A. Leutz
G9a-mediated Lysine Methylation Alters the Function of
Journal of Biological Chemistry (2008) 283 (39), 26357-26363
W.-H. Yang and D.B. Bloch
Probing the mRNA processing body using protein macroarrays
RNA (2007), 13:704-712.
Cepok S, Zhou D, Srivastava R, Nessler
S, Stei S, Büssow K, Sommer N, Hemmer B
Identification of Epstein-Barr virus proteins as putative
targets of the immune response in multiple
J Clin Invest. (2005) 115(5):1352-60
G. Grelle, S. Kostka, A. Otto, B.
Kersten, K. Genser, E. Müller, S. Wälter, A. Böddrich, U. Stelzl,
C. Hänig, R. Volkmer-Engert, C. Landgraf, S. Alberti, J.
Höhfeld, M. Strödicke, and E. Wanker
Identification of VCP/p97, CHIP and amphiphysin II
interaction partners using membrane-based human proteome
MCP (2005) 10.1074/mcp.M500198−MCP200
K. de Graaf, P. Hekerman, O. Spelten,
A. Herrmann, L. C. Packman, K. Bussow. G. Muller-Newen, W.
Characterisation of cyclin L2, a novel cyclin with an
arginine/serine-rich domain: phosphorylation by DYRK1A and
colocalisation with splicing factors.
J Biol Chem. (2004) 279(6):4612−24
K. Büssow, C. Quedenau, V. Sievert, J.
Tischler, C. Scheich, H. Seitz, B. Hieke, F.H. Niesen, F. Gotz, U.
Harttig, H. Lehrach
A catalog of human cDNA expression clones and its
application to structural genomics.
Genome Biol. 2004 Aug; 5(9):R71
U. Radelof, C.Hüls, B. Korn, J.
Proteinarrays und rekombinante Proteine für die
Laborwelt 2004. 5: p.35
Protein-Chips in der Genomforschung.
Laborwelt 2004. 4: p.12−15
J. Lee and M.T. Bedford
PABP1 identified as an arginine methyltransferase substrate
using high-density protein arrays.
EMBO Reports 2002. 3(3): p.268−73
U. Mahlknecht, O.G. Ottmann, and D.
Far-Western based protein-protein interaction screening of
high-density protein filter arrays.
Journal of Biotechnology 2001. 88(2): p. 89−94
K. Büssow, E. Nordhoff, C. Lübbert, H.
Lehrach, and G. Walter
A human cDNA library for high-throughput protein expression
Genomics 2000.65(1): 1−8
L. J. Holt, K. Büssow, G. Walter, and
I. M. Tomlinson
By-passing selection: direct screening for
using protein arrays.
Nucleic Acids Research 2000. 28(15): p. E72
K. Büssow, D. Cahill, W. Nietfeld, D.
Bancroft, E. Scherzinger, H. Lehrach, and G. Walter
A method for global protein expression and antibody
screening on high-density filters of an arrayed cDNA
Nucleic Acids Research 1998. 26(21): p. 5007−5008