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Target antibodies:
Our first-rate project management and purification strategies help guarantee a high success rate.
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Using polyclonal antibodies generated at Cambridge Research Biochemicals, the Sethi group at Cambridge University have identified a new phosphorylation site on the insulin receptor substrate 1 pleckstrin homology domain (IRS1-PH). Understanding insulin signalling pathways are crucial to the development of novel diagnostic and therapeutic strategies advancing the treatment of type II diabetes. Phosphorylation of insulin receptor substrate (IRS) proteins on serine residues is an important post-translational modification which is linked to insulin resistance. The generation of anti-pSer24-IRS1 was commissioned from Cambridge Research Biochemicals. Briefly, rabbit antisera were generated against the following N-terminally, keyhole limpet haemocyanin-conjugated phospho-peptide: i.e. [C]YLRKPK(p)SMHKRFF-acid. The phosphoreactive serum was affinity purified on a Thioprophyl Sepharose 6B column derivatised with nonphosphorylated antigen. The unbound antiserum was then passed down a Thiopropyl Sepharose 6B column derivatised with the phosphorylated peptide. The resulting triethylamine eluate retained significant phosphospecific immunoreactivity on peptide-coated ELISA plates and was used in Western blot analysis.
With thanks to Dr Jaswinder Sethi –(University of Cambridge) for granting permission to highlight her work. For further information please refer to : Sethi Lab
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Optimised Polyclonal Antibody Service:
CRB has been a provider of first-class antibodies to the pharmaceutical industry since 1985. We bring both a chemical and biological perspective to antigen design, while our first-rate project management and purification strategies help guarantee a high success rate.
Free Consultation:
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Expert advice on the vital first step of antigen prediction from either your target protein or peptide sequence |
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Antigens selected on the basis of antigenicity, protein structure and chemical suitability of peptide |
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Alignment with other proteins cross-checked to react or avoid specific sequence areas |
Superior Quality
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We use only high quality animals housed in UK Home Office, USDA & NIH compliant facilities |
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A variety of species and screening strategies employed for best approach |
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ISO9001:2008 Quality Accreditation |
Project Management
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Close involvement between CRB and client’s own goals to steer project |
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Fully-managed data from antibody generation programme |
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Online tracking facility available for easy access and storage of analytical data |
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Targeting of phosphorylation, glycosylation, sulphation, methylation and other PTM sites by the use of modified peptide antigens |
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Antigen design and purification strategies optimised to raise antibodies specific to neo-epitope or splice sites |
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Affinity purified sera is depleted against the non-modified peptide sequence to ensure specificity is obtained |
Validation & Labelling
At CRB we pride ourselves on the quality of our antibodies. We can now offer validation of our antibodies to confirm an anti-protein response.
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Immunoblotting (Western Blot) validation of polyclonal or monoclonal antibodies against target protein |
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Provision of Western Blot optimised protocol and working concentrations for custom antibody |
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Specificity against post translational modifications using positive and negative controls |
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Labelling of custom antibody with a range of enzymes, biotin or fluorescent dyes |
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Peptide Directed Antibodies
The use of peptide facilitates the production of antibodies for those occasions where a protein may not work such as:
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Post Translational Modification sites such as phosphorylation, glycosylation, sulphation, methylation, etc |
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Splice sites and neo-epitopes |
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Heterogeneous regions of conserved proteins |
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Cross-species antibodies |
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Transmembrane loops |
Carefully designed purification strategies make it possible to raise site-specific antibodies. Sera is first depleted against the non-modified sequence and then followed by antigen specific affinity purification to ensure specificity is obtained. |
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Purification of anti-phosphoresidue antibodies |
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Phospho Serine specific polyclonal antibody (anti-eIF2Βα and γ)
eIF5 has GDI activity necessary for translational control by eIF2 phosphorylation.
M.D Jennings & G.D. Pavitt - University of Manchester
Nature 2010, 465:378
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Phospho Serine specific polyclonal antibody (anti-pSer24-IRS1 [C]YLRKPK-(pS)-MHKRFF)
Regulation of Insulin Receptor Substrate 1 Pleckstrin Homology Domain By Protein Kinase C : Role of Serine 24 Phosphorylation.
R. Nawaratne, A. Gray, C. Jørgensen, C. P. Downes, K. Siddle, & J. K. Sethi - University of Cambridge & University of Dundee
Molecular Endocrinology 2006, 20 (8): 1838-1852
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Linear peptide antibody (antiG72 variant AY138546 [C]VTRKEGWKRRHEDGY-acid)
Investigation of G72 (DAOA) expression in the human brain.
I. Benzel, J. N. C. Kew, R. Viknaraja, F. Kelly, J. de Belleroche, S. Hirsch, T. H Sanderson, & P. R. Maycox – GSK & Imperial College London
BMC Psychiatry 2008, 8:94
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Anti-protein customer-supplied antibody (anti–IrrA)
Heme-responsive DNA Binding by the Global Iron Regulator Irr from Rhizobium leguminosarum.
C. Singleton, G. F. White, J. D. Todd, S. J. Marritt, M. R. Cheesman, A. W. B. Johnston & N. E. Le Brun - University of East Anglia
Journal of Biological Chemistry 2010, 285:16023-16031
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Should you require a quotation from any of our custom services, we have a clear cut and easy system that enables you to do so. Follow these three simple steps:
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Simply click on the relevant tab and register your details. |
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Once issued with a password you can then log on to our quotation system using your customer account log-in and a quotation will be issued within 24 hours. |
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At Cambridge Research Biochemicals we have a fast-track quotation system that tailors your requirements, gives you all the information you need and a specific price within 24 hours. You are also given a personal contact so that if you have any further questions you can directly contact one of our approachable representatives who has the technical knowledge to assist you.
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T+44 (0)1642 567180 F +44 (0)1642 567181 E enquiries@crbdiscovery.com
17-18 Belasis Court, Belasis Hall Technology Park, Billingham, Cleveland, TS23 4AZ United Kingdom
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