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ANTIBODIES  

                                                                  (anti-Human and others as indicated)

RDI Division of Fitzgerald Industries Intl  offers a wide line of  antibodies. Since no one antibody works best for all applications (neutralization, blotting, histochemistry, ELISA, etc), we offer many different types of antibodies to help solve this problem. Please inquire for other applications or types of antibodies not listed below.


Prion Antibodies ( monoclonal)
see also polyclonal anti-Prion

see m1839-ad.pdf (ad brochure)

see m1839-spec.pdf (unconjugated)     m1840.pdf  (Biotin conjugated)      m1841.pdf (HRP conjugated)

available:

unconjugated:            cat#RDI-M1839clb     $440.00/vial 250ul (0.5mg/ml)

Biotin conjugated       cat#RDI-M1840clb    $562.00

HRP conjugated         cat#RDI-M1841clb     $562.00

Immunogen CGGQWNKPSKPKTN (bovine PrP108-119 )

Isotype Mouse IgG1 K

A monoclonal antibody with high affinfity for proteinase K digested PrP Sc McAb 1E4 has its epitope at position 108-119 of the bovine prion protein. In contrast to many other antibodies 1E4 has a very broad species reactivity. This enables the detection of prion proteins in biological samples of many species including mouse adapted BSE (301V)-infected mice, scrapie-infected sheep, scrapie infected hamster (263K), CWD infected deer, sCJD- and vCJD-infected human on Western blots. The special feature of McAb 1E4 is that its epitope is mostly hidden on non-digested prion proteins, but after Proteinase K digestion the epitope becomes better available on protease resistant PrP Sc which results in a significant signal increase.

Applications: Western blot, ELISA, RIA, dot-blot, EliBlot*, FACS, immunohistochemistry.

* EliBlot, a new prion detection technique developed by Sanquin Research (van Oers et al 2005, submitted)

1E4 allows prion research in many immunological techniques across a broad range of species


Background information

The term “prion” was introduced by Stanley Prusiner in 1982 to describe the atypical infectious agent that causes transmissible spongiform encephalopathies, a group of infectious neurodegenerative diseases that include scrapie in sheep, Creutzfeldt-Jakob disease in humans, chronic wasting disease (CWD) in deer, and bovine spongiform encephalopathy (BSE) in cattle.

The ‘mad cow’ crisis has drawn a lot of attention to prion diseases. Significant progress has been made in prion disease research, and many aspects of prion pathogenesis are now understood. And yet the diagnostic procedures available for prion diseases are not nearly as sensitive as they ought to be, i.e. a diagnostic test to proof the presence of pathogenic prion in blood or serum.

At the moment there are a number of post-mortem tests on brain homogenates commercially available that have been found valid (100% sensitive and specific) even in symptomatic animals when testing cattle under the European Commission (E.g. Prionics, Abbott, BioRad, Inpro).

Still a lot of research and development is carried out in the field of prion proteins and this is the environment where 1E4 will be used.

Features of 1E4

In contrast to many other anti-prion antibodies, McAb 1E4 has a broad species reactivity (see attached leaflet). This enables the detection of prion proteins in biological samples of many species including mouse adapted BSE (301V)-infected mice, scrapie-infected sheep, scrapie infected hamster (263K), CWD infected deer, sCJD- and vCJD-infected human on Western blots.

1E4 has been tested in a broad variety of methods, such as Western blot, RIA, ELISA, EliBlot, FACS and immunohistochemistry.

1E4 allows prion research in many immunological

techniques across a broad range of species.

Most of the currently available TSE tests are based on the fact that PrPC, normal prion protein, is digested by Protease K, whereas PrPSc, TSE specific prion, is relatively resistant to degradation by proteases.

The special feature of McAb 1E4 is that the PrPSc epitope whereto it binds, PrP27-30, is mostly hidden on non-digested prion proteins, but after Proteinase K digestion the epitope becomes better available on proteinase resistant PrPSc which results in a significant signal increase.

After digestion with Protease K , McAb 1E4 binds to PrPSc with a high affinity, whereas it has a low affinity for non-digested PrPSc. This makes McAb 1E4 a highly interesting antibody in current prion disease research.


Literature references.

1. Wadsforth J D F et al., Tissue distribution of protease resistant prion protein in variant Creutzfeldt-Jakob disease using a highly sensitive immunoblotting assay (2001). Lancet 358, 171-180.

2. Schreuder B E et al., Preclinical test for prion diseases (1996). Nature 381, 563.

3. Naslavsky N, et al., Characterization of detergent-insoluble complexes containing the cellular prion protein and its scrapie isoform (1997). JBC 272, 6324-6331.

4. Wiessmann C, The ninth Datta Lecture. Molecular biology of transmissible spongiform encepholpathies (1996). FEBS Letters 389, 3-11.

5. Safar J et al., Eight prions strains have PrPSc molecules with different conformations (1998). Nature Medicine 4, 1157-1165.

6. Schmerr M J and Jenny A, A diagnostic test for scrapie-infected sheep using a capillary electrophoresis immunoassay with fluorescent-labelled peptides (1998). Electrophoresis 19, 409-413.

7. Houston F et al. Transmission of BSE by blood transfusion in sheep (2000). Lancet 356, 999-1000

8. Shaked G M et al., A protease-resistant protein isoform is present in urine of animals and humans affected with prion diseases (2001). J.Biol Chem 276 (34), 1479-82.

9. Kascsak R J et al., Mouse polyclonal and monoclonal antibody to scrapie-associated fibrin proteins (1987). J Virol 61, 3688-3693.

10. Laffling A J, et al. A monoclonal antibody that enables specific immunohistological detection of prion protein in bovine spongiform encephalopathy cases (2001). Neurosci Lett. 300, 99-102.

11. Korth C et al., Prion (PrPSc)-specific epitope defined by a monoclonal antibody (1997). Nature 390, 74-77.

12. MacGregor I, Prion protein and developments in its detection (2001). Transf Med 11, 3-14.

For In Vitro Research Use Only


Mouse (monoclonal) anti-Prion

catalog# RD-M1839clb     $438.00

Clone 1E4

This clone was derived from hybridization of SP2/0-Ag14 myeloma cells with spleen cells of a Prnp 0/0 mouse immunized with the peptide GQWNKPSKPKTN#(corresponding to the bovine PrP AA sequence 108-119; #=amidated carboxy-terminus).

Isotype IgG1 K

Source Culture supernatant

Purification Protein A affinity chromatography

Packing Each vial contains 250 µl (conc. 0.5 mg/ml) in PBS.

Preservative Merthiolate (0.001%)

Storage and stability Storage at -18°C to -32°C is recommended. Do not freeze and thaw more than three times. The reagent is guaranteed to remain stable until the expiry date stated on the vial label.

Major reactivity Monoclonal antibody 1E4 was isolated from hybridoma’s generated from spleen cells of a Prnp 0/0 mouse, immunized with peptide GQWNKPSKPKTN# (corresponding to the bovine PrP aminoacid sequence 108-119; #=amidated carboxy-terminus) coupled to KLH at its N-terminal end via a CG-AA linker. The clone was selected due to its specific binding behaviour; on Western blot a strong binding reaction was found to BSE brain homogenates digested with Proteinase K. This was in contrast with a weak binding to undigested BSE brain homogenates, suggesting that 1E4 has a higher affinity for Proteinase K cleaved PrP 27-30 than for the non-cleaved PrP Sc . Furthermore the Western blot also revealed a weak binding onto non-digested brain homogenate from a normal cow. This is in contrast with other commercially available antibodies, most of which express a similar affinity for both PrP conformers and cleaved PrP 27-30 . Beside BSE infected cattle, MAb 1E4 also reacted with prions from mouse adapted BSE (301V)-infected mice, scrapie-infected sheep, scrapie infected hamster (263K), CWD infected deer, sCJD- and vCJD-infected human on Western blots. However the striking difference between the affinity for cleaved and non-cleaved PrP Sc observed for BSE in cattle is not observed in these samples. Molecular mass The molecular weight of both PrP C and PrP Sc is 30-35 kD; after digestion with protease, PrP Sc becomes PrP 27-30 (27-30 kD).

Application Prion research on biological samples, body fluids, cells, tissue sections and homogenates, capturing or detecting antibody in immunoassays

Methods Western blot, RIA, ELISA, EliBlot, FACS, immunohistochemistry.


Mouse (monoclonal) anti-Prion, Biotin

Cat#RDI-M1840clb    $562.00

Clone 1E4

This clone was derived from hybridization of SP2/0-Ag14 myeloma cells with spleen cells of a Prnp 0/0 mouse immunized with the peptide GQWNKPSKPKTN#(corresponding to the bovine PrP AA sequence 108-119; #=amidated carboxy-terminus).

Isotype IgG1 K

Source Culture supernatant

Purification Protein A affinity chromatography

Conjugation The monoclonal antibodies were conjugated to biotin using a modification of the procedure according to Hnatowich. Packing Each vial contains 250 µl (conc. 0.5 mg/ml) in 20 mM TRIS, 150 mM NaCl and 1% BSA.

Preservative Merthiolate (0.001%)

Storage and stability Storage at -18°C to -32°C is recommended. Do not freeze and thaw more than three times. The reagent is guaranteed to remain stable until the expiry date stated on the vial label.

Major reactivity Monoclonal antibody 1E4 was isolated from hybridoma’s generated from spleen cells of a Prnp 0/0 mouse, immunized with peptide GQWNKPSKPKTN# (corresponding to the bovine PrP aminoacid sequence 108-119; #=amidated carboxy-terminus) coupled to KLH at its N-terminal end via a CG-AA linker. The clone was selected due to its specific binding behaviour; on Western blot a strong binding reaction was found to BSE brain homogenates digested with Proteinase K. This was in contrast with a weak binding to undigested BSE brain homogenates, suggesting that 1E4 has a higher affinity for Proteinase K cleaved PrP 27-30 than for the non-cleaved PrP Sc . Furthermore the Western blot also revealed a weak binding onto non-digested brain homogenate from a normal cow. This is in contrast with other commercially available antibodies, most of which express a similar affinity for both PrP conformers and cleaved PrP 27-30 Beside BSE infected cattle, MAb 1E4 also reacted with prions from mouse adapted BSE (301V)-infected mice, scrapie-infected sheep, scrapie infected hamster (263K), CWD infected deer, sCJD- and vCJD-infected human on Western blots. However the striking difference between the affinity for cleaved and non-cleaved PrP Sc observed for BSE in cattle is not observed in these samples. Molecular mass The molecular weight of both PrP C and PrP Sc is 30-35 kD; after digestion with protease, PrP Sc becomes PrP 27-30 (27-30 kD).

Application Prion research on biological samples, body fluids, cells, tissue sections and homogenates, capturing or detecting antibody in immunoassays

Methods Western blot, RIA, ELISA, EliBlot, FACS, immunohistochemistry.


Mouse (monoclonal) anti-Prion, HRP conjugated

Cat.no   RDI-M1841clb    $562.00

Clone 1E4

This clone was derived from hybridization of SP2/0-Ag14 myeloma cells with spleen cells of a Prnp 0/0 mouse immunized with the peptide GQWNKPSKPKTN#(corresponding to the bovine PrP AA sequence 108-119; #=amidated carboxy-terminus).

Isotype IgG1 K

Source Culture supernatant

Purification Protein A affinity chromatography

Conjugation The monoclonal antibodies were conjugated to HRP by a modified way of the procedure according to Wilson and Nakane.

Packing Each vial contains 250 µl (conc. 0.5 mg/ml) in 20 mM TRIS, 150 mM NaCl and 1% BSA.

Preservative Merthiolate (0.001%)

Storage and stability Storage at -18°C to -32°C is recommended. Do not freeze and thaw more than three times. The reagent is guaranteed to remain stable until the expiry date stated on the vial label.

Major reactivity Monoclonal antibody 1E4 was isolated from hybridoma’s generated from spleen cells of a Prnp 0/0 mouse, immunized with peptide GQWNKPSKPKTN# (corresponding to the bovine PrP aminoacid sequence 108-119; #=amidated carboxy-terminus) coupled to KLH at its N-terminal end via a CG-AA linker. The clone was selected due to its specific binding behaviour; on Western blot a strong binding reaction was found to BSE brain homogenates digested with Proteinase K. This was in contrast with a weak binding to undigested BSE brain homogenates, suggesting that 1E4 has a higher affinity for Proteinase K cleaved PrP 27-30 than for the non-cleaved PrP Sc . Furthermore the Western blot also revealed a weak binding onto non-digested brain homogenate from a normal cow. This is in contrast with other commercially available antibodies, most of which express a similar affinity for both PrP conformers and cleaved PrP 27-30 . Beside BSE infected cattle, MAb 1E4 also reacted with prions from mouse adapted BSE (301V)-infected mice, scrapie-infected sheep, scrapie infected hamster (263K), CWD infected deer, sCJD- and vCJD-infected human on Western blots. However the striking difference between the affinity for cleaved and non-cleaved PrP Sc observed for BSE in cattle is not observed in these samples. Molecular mass The molecular weight of both PrP C and PrP Sc is 30-35 kD; after digestion with protease, PrP Sc becomes PrP 27-30 (27-30 kD).

Application Prion research on biological samples, body fluids, cells, tissue sections and homogenates, capturing or detecting antibody in immunoassays

Methods Western blot, RIA, ELISA, EliBlot, immunohistochemistry.

For In Vitro Research Use Only


 RDI Division of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

phone (978) 371-6446 or (800) 370-2222

fax     (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

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