anti-bovine prion protein, biotinylated
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 PBS/Casein-buffer pH 8.0 with 0.01% thimerosal as preservative)
Monoclonal antibody (mAb) 5G12, a murine IgG1K subtype antibody, purified by affinity chromatography and conjugated with biotin.
mAb 5G12 biotinylated may be used for Western and dot blots at a dilution of 1:500-1:2000 and in ELISA at 1:1000-10000.
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, and faintly with that from mouse and hamster. It detects the native bovine PrPC and PrPSc in dot blots, the partly refolded recombinant bovine PrPC in ELISA applications, and denatured PrP in immunoblots of brain homogenates of healthy and BSE-affected animals.
A) PrP from brain homogenates of healthy cattle (lane 1) appears with three glycosylation-dependent bands on Western blots. Treatment of the homogenates of BSE-affected animals with proteinase K under defined conditions degrades the abundant PrPP molecules, revealing the shortened PrPSc (lane 2 without dilution of a 10 % homogenate (w/v), 3-1:10, 4-1:50 and 5-1:100). 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 (lane 6-50 ng, 7-5 ng and 8-0.5 ng).
B) Recombinant bovine PrPC and PrP from brain homogenates of healthy cattle was analyzed in an ELISA sandwich test using microtiter plates coated with mAb 3C10 (JenaBioscience) and mAb 5G12biotinyl as detection antibody.
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.