anti-bovine prion protein
murine, monoclonal, clone 5G12
For in vitro use only!
Shipping: shipped on dry ice
Storage Conditions: store at -80 °C
avoid freeze/thaw cycles
Shelf Life: 12 months
Form: liquid (Supplied in culture medium supplemented with FBS)
Monoclonal antibody (mAb) 5G12 is a murine IgG1K subtype antibody.
mAb 5G12 may be used for species-specific Western blotting, dot blots and ELISA at a dilution of 1:10000 - 1:30000.
mAb 5G12 recognizes a sequence containing Y236YQR239 (human PrP: amino acids 225-228) in the C-terminal part of the prion protein. mAb 5G12 reacts strongly with recombinant PrPc from cattle, sheep and human in Western blots, but only faintly with that from mouse and hamster. It detects the native bovine PrPC and PrPSc in dot blots, the partly refolded recombinant bovine PrP in ELISA applications, and the denatured form in immunoblots of brain homogenates of healthy and BSE-affected animals.
PrP from brain homogenates of healthy cattle (lanes 1 and 4) appears with three glycosylation-dependent bands, with masses between 16 and 36 kDa, on Western blots. Treatment of the homogenates of BSE-affected animals with proteinase K under defined conditions degrades the abundant PrPC molecules, revealing the shortened PrPSc (lanes 2 and 5). Proteinase K cleaves some residues from PrPSc, causing an apparent decrease in its molecular mass. Recombinant bovine PrPC runs at 27 kDa, the dimer at 48 kDa (lanes 3 and 6).
Species and prion protein sequence:
Human, Chimpanzee, Cattle, Goat, Sheep: YYQ---R
Hamster, Mouse, Rat: YYDGRR
Schaller et al. (1999) Validation of a western immunoblotting procedure for bovine PrPSc detection and its use as a rapid surveillance method for the diagnosis of bovine spongiform encephalopathy (BSE). Acta Neuropathologica 98:437.
Kübler et al. (2003) Diagnosis of prion diseases. Br. Med. Bull. 66:267.
Kim et al. (2004) Antigenic characterization of an abnormal isoform of prion protein using a new diverse panel of monoclonal antibodies. Virology 320:40.