Immunochemistry

Swedclone is concerned to ensure a successful and rapid result which fulfils the specific demands from our clients. Swedclone therefore offers a close collaboration where particular demands can be met. We routinously provide immunochemistry services such as Affinity Purification, Protein A/G Purification, dot blot and western analysis as well as ELISA to characterize and prepare custom antibodies for use in assays.

A pre-screen of the fraction within the polyclonal mouse-sera ensures a correct immune response prior to cell-fusion, which saves time and enhance rate of success.

Affinity Purification


Affinity purification is frequently used upon purification of the desired antibody from complex mixtures such as whole serum samples containing e.g. polyclonal serum samples and ascitis fluids (due to ethical reasons SwedClone does not perform expansion of hybridomas in ascitis fluid).
The procedure is based on covalent coupling of the antigen to an affinity column. Serum/cell culture media is then passed through the column, thereby binding antigen-specific antibodies and allowing non-specific antibodies to pass through.

  • Affinity Purification – 25 ml serum: 320 €
  • Affinity Purification – 25 ml egg yolk: 425 €
  • Affinity purification monoclonals 1000 ml enquire
  • Scale up enquire

IgG Purification


IgG purification using protein A/G and L provides an efficient method to separate immunoglobulin (antibodies) in complex mixtures, thereby reducing some background in assays.
However, please note that this will not provide specificity to the immunizing antigen and so affinity purification is the preferred technique for most applications.

  • IgG Purification – 25 ml serum: 180 €


IgY Purification


The terms IgG and IgY are commonly interchanged when speaking of chicken immunoglobulin. Immunoglobulins from chickens and other avian species bear some resemblance to mammailian IgG, but also display some unique structural and functional characteristics that distinguish them from IgG. IgY is the accepted/proper term for chicken antibodies. Chicken IgY is the functional equivalent to mammalian IgG. It is found in the serum of chickens and is passed from the mother chicken to the embryo via the egg yolk, imparting a high concentration of chicken IgY to developing embryo. The "Y" in IgY comes from "yolk" and is the main antibody component in the egg yolk. Protein L is typically used for IgY purifications.

  • IgY Purification – 25 ml egg yolk: 280 €

Western and dot blot analysis


The western blot (alternatively, immunoblot) is a method of detecting specific proteins in a given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide. The proteins are transferred to a membrane (typically nitrocellulose or PVDF), where they are probed (detected) using antibodies specific to the target protein.
Dot-blot uses a similar approach the protein is however in this case the antigen is not separated on through gel electrophoresis before transfer to membrane. Using pure antigen dot-blot is a powerful and rapid technique.

  • Western analysis 500 €
  • Dot blot analysis 280 €


ELISA


Enzyme linked immunosorbent assay (ELISA) is routinely used in the screening process of identifying and characterising positive hybridom clones. The antigen is bound to the surface of a polystyrene well, blocked to reduce non specific binding, incubated with various dilutions of the test serum and allowed to react with enzyme linked secondary antibody to the appropriate species being tested. The final stage shows colour development due to reaction between the enzyme conjugate and the appropriate substrate. SwedClone recommends running an ELISA on the first production bleed of peptide projects before proceeding with affinity purifications.

  • Always included
 
 
SwedClone | Uminova Science Park | Box 7964 | SE-907 19 Umeå, Sweden | Phone: +46 90 15 49 40 / +46 70 655 9440 | E-mail: info@swedclone.com
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