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An assay’s ability to specifically and sensitively detect a molecule of interest is critically based on suitable antigenic molecule design. Our scientists design peptides and organic derivatives to allow for the generation of high quality polyclonal and monoclonal antibodies as well as suitable enzyme and luminescent detection conjugates for immunoassay use.
- Under our direction, the high quality, chemical organic synthesis companies we work with make biological molecule derivatives suitable for effective conjugation.
- We carry out protein labeling with the derivatives or peptides, utilizing porcine thyroglobulin (PTG) for raising antibodies and BSA for ELISA testing.
- We prefer using PTG over KLH as a carrier protein because of its solubility and size. PTG can be dissolved in aqueous buffers at 20 mg/mL (far in excess of KLH) and is comprised of 4 subunits each of about 165kDa in size. This solubility can help in estimating incorporation ratios of antigen to PTG spectrophotometrically.
- For free thiol group containing derivatives, we pre-treat the PTG with a reagent to add maleimide groups.
- For those derivatives with a free carboxylic acid, we chemically activate using DCC/NHS for coupling to protein lysine groups.
- The PTG and BSA labeled antigens are purified on desalting columns, followed by analysis to estimate labeling efficiencies when possible.
- Please contact us at contracts [at] ArborAssays.com for our input into your project.
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