Research Biochemicals. ANTIBODIES TO PRION PROTEINS

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1 Research Biochemicals. ANTIBODIES TO PRION PROTEINS 2005_03

2 Research Biochemicals. TABLE OF CONTENTS MONOCLONAL AB 6H4 3 MONOCLONAL AB 34C9 4 RECOMBINANT BOVINE PRP 5 PRICE LIST 7 ORDER INFORMATION 8 2

3 Antibodies. MONOCLONAL ANTIBODY 6H4 Product No µg monoclonal anti-prp antibody 6H4 (mab 6H4) mg monoclonal anti-prp antibody 6H4 (mab 6H4) monoclonal antibody to prion protein Product description Monoclonal antibody (mab) 6H4 is a mouse IgG1 subtype antibody. Light chain: κ subtype Concentration Culture supernatant; approx mg/ml, see vial label. > 95% pure antibody Specificity mab 6H4 recognizes the sequence DYEDRYYRE in the prion protein (human PrP: amino acids ). This sequence is conserved in most known mammalian PrP sequences (human, cattle, sheep, rabbit, mink, and a variety of primates). In mouse, hamster and rat, the tyrosine at position 145 is replaced by a tryptophan. However, mab 6H4 reacts on Western blots with mouse and hamster PrP indicating that the substitution at position 145 does not prevent binding of 6H4. Stability and storage mab 6H4 was produced in serum-free medium. There are essentially no other proteins present. In the TM medium, phenol red and basic nutrients like amino acids are still present. The antibody is supplied with 10 mm sodium azide as preservative. For long term storage, the antibody can be kept frozen at -20 C. Repeated freezing/thawing cycles should be avoided. Applications mab 6H4 can be used for Western blotting and ELISA at a dilution of 1: : Immunohistochemical staining of PrP is currently performed in various laboratories. (see application literature 3

4 Antibodies. MONOCLONAL ANTIBODY 34C9 Product No µg monoclonal anti-prp antibody 34C9 (mab 34C9) mg monoclonal anti-prp antibody 34C9 (mab 34C9) monoclonal antibody to prion protein Product description Monoclonal antibody (mab) 34C9 is a mouse IgG2b subtype antibody. Light chain: κ subtype Concentration Culture supernatant; approx mg/ml, see vial label. > 95% pure antibody Specificity mab 34C9 recognizes the sequence LIHFG in the bovine prion protein (corresponding to position with numbering according to the human PrP). This sequence is conserved in bovine, rabbit, pig, and mule deer PrP sequences but not in human, mouse or hamster PrP. Stability and storage mab 34C9 was produced in serum-free medium. There are essentially no other proteins present. In the TM medium, phenol red and basic nutrients like amino acids are still present. The antibody is supplied with 10 mm sodium azide as preservative. For long term storage, the antibody can be kept frozen at -20 C. Repeated freezing/thawing cycles should be avoided. Applications mab 34C9 can be used for Western blotting at a dilution of 1: :

5 Recombinant Proteins. RECOMBINANT BOVINE PrP Product No µg lyophilized recombinant bovine PrP (rec bovprp) recombinant bovine prion protein Product description Highly purified recombinant bovine prion protein (rec bovprp) (Ref. 1). The nucleotide sequence corresponding to the mature primary structure of bovine PrP (amino acids ) was amplified by PCR from genomic DNA. The PCR product was cloned into pprio and expressed in an E. coli K12 strain. Expressed rec bovprp was solubilized in 6 M Guanidine Hydrochloride from inclusion bodies, oxidized, and purified by various chromatographic steps. The MW is Da. The extinction coefficient in aqueous solution at 280 nm is M -1 xcm -1. Sequence MKKRPKPGGGWNTGGSRYPGQGSPGGNRYPPQGGGGWGQPHGGGWGQPHG 50 GGWGQPHGGGWGQPHGGGWGQPHGGGGWGQGGTHGQWNKPSKPKTNMKHV 100 AGAAAAGAVVGGLGGYMLGSAMSRPLIHFGSDYEDRYYRENMHRYPNQVY 150 YRPVDQYSNQNNFVHDCVNITVKEHTVTTTTKGENFTETDIKMMERVVEQ 200 MCITQYQRESQAYYQRGAS 219 Stabilizer & Preservatives None Stability and storage Lyophilized: Stable at -20 C if not opened. Lyophilized protein is dissolved best in H 2 O. Dissolving in a buffer might lead to precipitation. Applications Recombinant bovine PrP is ideal as standard and positive control for Western Blot and ELISA. Hazard Data Recombinant PrP of any species has to date not been shown to contain infectivity (Ref. 2-4). However, it should be noted that this does not conclusively exclude possible hazardous effects under all circumstances, particularly since the details of the molecular mechanism underlying the propagation of the TSE infectious agent have not yet been resolved (Ref. 5-7). 5

6 Handling and Safety Guidelines Handle according to local legal guidelines. Avoid contact with eyes, skin, and clothing. Avoid aerosol forming procedures. Do not ingest. References 1 Korth C, Stierli B, Streit P, Moser M, Schaller O, Fischer R, Schulzschaeffer W, Kretzschmar H, Raeber A, Braun U, Ehrensperger F, Hornemann S, Glockshuber R, Riek R, Billeter M, Wuthrich K, Oesch B (1997). Prion (PrP Sc )-specific epitope defined by a monoclonal antibody. Nature 390 (6655), Hill AF, Antoniou M, Collinge J (1999). Protease-resistant prion protein produced in vitro lacks detectable infectivity. J Gen Virol 80, Post K, Brown DR, Groschup M, Kretzschmar HA, Riesner D (2000). Neurotoxicity but not infectivity of prion proteins can be induced reversibly in vitro. Arch Virol Suppl 2000 (16), Volkel D, Blankenfeldt W, Schomburg D (1998). Large-scale production, purification and refolding of the full-length cellular prion protein from Syrian golden hamster in Escherichia coli using the glutathione S-transferase-fusion system. Eur J Biochem 251(1-2), Kaneko K, Petetz D, Pan KM, Blochberger TC, Wille H, Gabizon R, Griffith OH, Cohen FE, Baldwin MA, Prusiner SB (1995). Prion Protein (PrP) synthetic peptides induce cellular PrP to acquire properties of the scrapie isoform. Proc Natl Acad Sci USA 92, Weissmann C (1999). Molecular genetics of transmissible spongiform encephalopathies. J Biol Chem 274(1), Prusiner SB (1998). Prions. Proc Natl Acad Sci U S A 95(23),

7 Research Biochemicals. TRANSFER PRICE LIST 2005 Product Number Product Name Pack Size Quantity Price (in CHF) Price (in ) Price (in USD) Antibodies µ mab 6H4 100 g 1 1' mab 6H4 1 mg 1 8' ' '800.- µ mab 34C9 100 g 1 1' mab 34C9 1 mg 1 8' ' '800.- rec PrP µ rec PrP bovine 100 g

8 Research Biochemicals. ORDER INFORMATION On request we will be pleased to send you the general terms and conditions Price Rebate Order Address DDU (transport costs will be charged separately) 10% for Universities (for Antibodies and/or rec PrP) Prionics AG Order Intake Wagistrasse 27a CH-8952 Schlieren-Zurich Switzerland PHONE: FAX:

9 8952 SCHLIEREN

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