Antibodies were prepared by injecting laying hens with purified recombinant human PRN protein. After a series of boosts, eggs were collected from hyperimmunized aniamsl and the IgY fraction prepared. Affinity purified antibodies were then isolated from the IgY fraction using a PRN protein column. Finally, affinity purified antibodies were mixed with the IgY fraction to make the final product. These antibodies have been validated with human, mouse and rat tissues.
Mouse fibroblast cultures transfected with a recombinant fragment of the mouse prion protein (green staining). Non-infected cells show labeled nuclei (blue staining) due to treatment with Hoechst stain.
Prion protein is a ubiquitously-expressed, 27661 dalton protein (253 amino acids, NP_898902.1) of unclear function(s) in eukaryotes. This protein can exist in one of two conformational forms. One is its normally-folded form (PrPc or "cellular PrP"), which contains a mixture of alpha helical regions and a few beta-pleated sheets. The other is a misfolded form (PrPsc or "scrapie PrP") whose structure contains mostly beta-pleated sheets. This misfolded scrapie PrP form is cytotoxic, especially to nervous tissue, and can serve as a "seed," inducing adjacent, normally-folded PrP proteins to misfold into the cytotoxic conformation. In this sense, the scrapie PrP can be thought of as an infectious agent both within the organism as well between organisms.
Mixture of IgY fraction and affinity-purified antibodies
10 mg/mL
Polyclonal
IgY
ICC, IHC, WB
Chicken
PRNP ALTPRP PRIP PRP
27 kDa
Reccombinant fragment of human prion
Human, Mouse, Rat
AB_2313559
Store at -20°C in the dark. Under these conditions, the antibodies should have a shelf life of at least twelve months, provided they remain sterile.
Liquid
Chickens were immunized with a recombinant fragment of the human prion protein emulsified with Freund’s complete and incomplete adjuvants.After multiple injections, eggs were collected from the hens, and IgY fractions were prepared from the yolks.Affinity-purified antibodies were then prepared using an agarose column to which the recombinant protein fragments were covalently attached. Finally, affinity purified antibodies (50 µg/mL final concentration) were mixed with the IgY fraction (10 mg/mL final concentration) in a mixture containing glycerol (50% v/v) (to prevent freezing at -20°C) and 0.02% sodium azide.
Sodium phosphate (10 mM, pH 7.2) buffered isotonic saline (0.9%, w/v), glycerol (50%, v/v), with sodium azide (0.02%,w/v) as an anti-microbial agent.
