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CRB Discovery

Case Study: Phospho-Specific Antibody

Phosphoserine specific polyclonal antibody
anti-pSer24-IRS1 using phospho peptide

Target antibodies:

Our first-rate project management and purification strategies help guarantee a high success rate.

The Key to Success:

Affinity Purification strategy to deliver phospho-specific antibodies

Target Antibodies
This service provides the highest quality of target specific polyclonal antibodies that we offer
Find out more

Anti-pSer24 - IRS1: Lipid binding and intracellular 
localisation of the IRS1-PH domain
Western blot analysis showing PMA-induced phosphorylation of Ser24, suggesting that calcium-dependent PKCs are activated by PMA and are required to induce phosphorylation of IRS-1. IRS-1 Serine 24

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

Read more Find out more about our Antibody Production
Target Antibodies

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:


Expert advice on the vital first step of antigen prediction from either your target protein or peptide sequence


Antigens selected on the basis of antigenicity, protein structure and chemical suitability of peptide


Alignment with other proteins cross-checked to react or avoid specific sequence areas

Superior Quality


We use only high quality animals housed in UK Home Office, USDA & NIH compliant facilities


A variety of species and screening strategies employed for best approach


ISO9001:2008 Quality Accreditation

Project Management


Close involvement between CRB and client’s own goals to steer project


Fully-managed data from antibody generation programme


Online tracking facility available for easy access and storage of analytical data

Protein Modifications


Targeting of phosphorylation, glycosylation, sulphation, methylation and other PTM sites by the use of modified peptide antigens


Antigen design and purification strategies optimised to raise antibodies specific to neo-epitope or splice sites


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.


Immunoblotting (Western Blot) validation of polyclonal or monoclonal antibodies against target protein


Provision of Western Blot optimised protocol and working concentrations for custom antibody


Specificity against post translational modifications using positive and negative controls


Labelling of custom antibody with a range of enzymes, biotin or fluorescent dyes

Download our Antibody Buyer’s Guide
More about antibody production
Phospho-specific Antibodies Take a look at our Antibody Production section on our website to find out more

Peptide Directed Antibodies

The use of peptide facilitates the production of antibodies for those occasions where a protein may not work such as:


Post Translational Modification sites such as phosphorylation, glycosylation, sulphation, methylation, etc


Splice sites and neo-epitopes


Heterogeneous regions of conserved proteins


Cross-species antibodies


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.

Purification of anti-phosphoresidue antibodies

Purification of anti-phosphoresidue antibodies

To order your polyclonal antibodies visit our website
or telephone: 01642 567180
Download our Antibody Buyer's Guide
Download our Antibody Buyer's Guide

Case Studies: Customer References

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

Request a Quotation

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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Go to:

Simply click on the relevant tab and register your details.

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.

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.

Meet a CRB representitive

If you would like any support, help or technical advice we would like to hear from you.

CRB is delighted to be sponsoring the Cambridge Cell Biology Seminar Series and the next event is being held on the 24th March 2011.
Dr Melina Schuh-MRC- Laboratory of Molecular Biology, Cambridge:
Meiosis in mammals-expanding the view
Adhesive control of cell polarity, organelle positioning and cell division axis in growing cells and of primary ciliogenesis in cell-cycle arrested cells.
24th March 4pm Sackler Lecture Theatre Lent Term University of Cambridge
Click here to find out more
CRB will be exhibiting and attending the following exhibitions:
Peptide Arrays Meeting AACR
BioTrinity 2011 Signalling Meeting

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F +44 (0)1642 567181

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