The Thermo Scientific Pierce Chicken IgY Purification Kit purifies chicken antibodies from hen egg yolks using a unique delipidation reagent to separate protein from lipid and a precipitation reagent to selectively isolate the IgY fraction.
Typical IgG affinity resins (Protein A or Protein G) do not bind chicken IgY and therefore are not effective for IgY purification. The Pierce Chicken IgY Purification Kit uses a delipidation-and-precipitation procedure that is optimized to recover approximately 100mg of 90% pure IgY from fresh egg yolks of appropriately immunized chickens. The resulting polyclonal IgY is sufficiently pure for direct use in many immunodetection procedures, or it can be easily affinity-purified against specific antigen because it is dissolved in PBS and thoroughly clarified.
- More for your money – purifies twice the amount of IgY than the leading competitor’s kit with a lower cost-per-mg of IgY purified
- High recovery and purity – obtain 80 to 120mg IgY per egg yolk at 85% to 95% purity by SDS-PAGE analysis
- Ease-of-use – simple precipitation method purifies chicken IgY without affinity columns
- Trouble-free method – fewer steps than other IgY purification kits
- Flexibility – eggs can be stored in buffer and purified at a later date
- Convenience – use eggs directly out of refrigerator; no need to wait for them to warm up
Affinity chromatography with Protein A or Protein G cannot be used to purify IgY from chicken serum or egg yolk because this species of immunoglobulins are not recognized by the Protein A, G or L binding sites. However, the Pierce Chicken IgY Purification Kit provides a simple and reliable method for recovering large amounts of chicken IgY from egg yolk with our unique delipidation reagent that separates the proteins from lipid. (The egg yolk can be stored in the delipidation reagent a year or longer before processing.) After lipid separation, simply mix the sample with the IgY Precipitation Reagent which causes the IgY to separate from the lipid fraction during centrifugation. The precipitated chicken IgY is then easily recovered by dissolution in phosphate-buffered saline (PBS).
|Protocol summary for the Thermo Scientific Pierce IgY Purification Kit. Yolks and whites are separated (1). Delipidation reagent is added (2), followed by centrifugation (3) and separation from lipid solids. Then, IgY precipitation reagent is added (4), followed by centrifugation to pellet the IgY (5). After the liquid is decanted, the purified IgY is dissolved in phosphate-buffered saline (6).
||Better IgY Purification yield and purity with Thermo Scientific Pierce Chicken IgY Purification Kit. Non-reducing SDS-PAGE analysis of IgY samples. Compared to a pure IgY control sample (Lane 1), results indicate that the IgY purified with Supplier Z Kit was significantly degraded (see bands at 75, 50 and 25K). By contrast the Pierce Chicken IgY Purification Kit gave a high yield of very pure whole IgY (only prominent band is at 150 kDa).
Chicken IgY vs. Mammalian IgG:
The terms IgG and IgY are commonly interchanged when referring to chicken immunoglobulin. Immunoglobulins from chickens and other avian species bear some resemblance to mammalian 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.
Advantages of Chicken IgY:
Chicken IgY immunoglobulins can be harvested from the egg yolk as well as serum and blood. There are many advantages to using these antibodies:
- Unlike mice, rabbits or other mammals, hens elicit a strong antibody response against highly conserved mammalian protein sequences
- Chicken antibodies can be obtained without sacrificing or bleeding the animal – simply collect the eggs and use the Pierce Chicken IgY Purification Kit
- Each yolk can yield 100-150mg of antibody.
- Chicken IgY has less cross-reactivity toward mammalian proteins – does not bind Fc receptors!
- Akita, E.M., et al. (1993). J. Immunol. Methods 160, 207.
- Jensenius, J.C., et al. (1981). J. Immunol. Methods 46, 63.
- Larsson, A., et al. (1993). Poultry Sci. 72, 1807.
- Larsson, A., et al. (1990). Comp. Immun. Microbiol. Infect. 13, 199.
- Polson, A., et al. (1985). Immunol. Invest. 14, 323.
- Cassidy, P.B., et al. (2006). Carcinogenesis. 27, 2538-2549.
- Kamiya, Y., et al. (2005).J. Biol. Chem. 280, 37178-37182.
- Kantardzhieva, A. et al. (2005). Retinal Cell Biol. 46, 2192-2201.
- Jayasimha Rao, J., et al. (2003) Biol. Reprod.68, 290-301.
- Sommerville, J., et al. (2005) J. Cell Sci.118, 1299-1307.
Pierce Chicken IgY Purification Kit FAQ
Review of antibody purification methods
Anti-Chicken Secondary Antibodies
Goat Anti-IgY Antibody, Unlabeled (PA1-28653)
Goat Anti-IgY Antibody, Alkaline Phosphatase Conjugated (PA1-30369)
Goat Anti-IgY Antibody, Horseradish Peroxidase Conjugated (PA1-28658)
Anti-IgY Agarose Resin (SA1-9588)