Developing Custom Rabbit Monoclonal Antibodies Against Small Molecule Haptens

Raising Custom Monoclonal Antibodies in Rabbits Against Small Molecules

Small polymers and molecules are found with extreme frequency in medicines, foods, cosmetics, narcotics, pesticides and research reagents. These are also common in the environment and human body.

This antigen category requires to be probed, for which reason antibodies are being raised against them for a rising number of applications:

  • Disease diagnostics, prognosis and therapy
  • Monitoring therapy
  • Detecting and quantifying narcotics and drugs
  • Monitoring environmental substances

It is known that it is easy to raise a brisk immune response against a variety of small molecule haptens in rabbits, if the small molecules are properly conjugated to larger carrier proteins that can provoke an immune reaction.

Generation of specific antibodies against small molecule haptens.

Figure 1 - Generation of specific antibodies against small molecule haptens.

Abcam has its RabMAb® platform that allows monoclonal antibodies of high affinity and specificity to be created and chosen to suit each application separately.

Some instances of matching antibodies to project applications:

Anti-Thiophosphate Ester RabMAb Primary Antibody (ab92570) ​

This antibody was developed in association with Dr. Kevan Shokat, a UCSF researcher, to be used when generating a profile for protein kinase substrates. It allows:

  • Thiophosphorylated proteins to be detected specifically
  • Recognition of all residues which have undergone Ser-, Thr-, Tyr-, His-, and Asp-thiophosphorylation in any context
  • Unique specificity, distinguishing thiophosphate alkylation from other modes of alkylation, in contrast to chick antibodies
  • Replacement of radioactive labeling assay with another method which is easy and safe, besides being ecologically sound

View the Anti-Thiophosphate ester RabMAb primary antibody [51-8] (ab92570) datasheet

Western blot analysis using Thiophosphate Ester Specific RabMAb primary antibody (ab92570)*. In vitro kinase reaction using JNK1 protein (10ng) incubated with c-jun-GST substrate (1ug) using ATP-gamma-S followed by alkylation with PNBM (2.5mM).

In the above reaction, the kinase phase was performed at room temperature for 30 minutes. PNBM alkylation was then carried out at room temperature for 2 hours, the concentration being 2.5 mM.

The products of the reaction were examined using western blotting. The antibody used was thiophosphate ester specific RabMAb primary antibody (ab92570) at 1:5000. The verification of equal loading was done using a-GST antibody. It should be remembered that PNBM reacts with cysteine residues as well, thus lowering the affinity with which other antibodies and antigens react.

Anti-Progesterone RabMAb Primary Antibody (PROG-124)

This antibody is an anti-Progesterone RabMAb primary which was developed in collaboration with Pantex, a division of Bio-Analysis. It has the following features:

  • Very high sensitivity of 0.03nM as well as selectivity
  • Shows linear interactions over its dynamic range
  • Produces results that are consistent with those published using patient test samples  

The following chart shows its specific binding with progesterone rather than other steroid hormones.

About Abcam

Abcam is a global life sciences company providing highly validated antibodies and other binders and assays to the research and clinical communities to help advance the understanding of biology and causes of disease.

Abcam’s mission is to serve life scientists to help them achieve their mission faster by listening to their needs, continuously innovating and improving and by giving them the tools, data and experience they want. Abcam’s ambition is to become the most influential life science company for researchers worldwide.

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Last updated: Jun 6, 2019 at 9:49 AM


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