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Specifications
| Product Type: | Antibody |
| Name: | GP25-200/P23 Antibodies |
| Antigen: | GP25-200 or P23 |
| Molecular Weight: | Multiple 25- to >200-kDa (glyco) proteins (GP25-200); 23-kDa protein (P23) |
| Isotype: | See Comments section below for details on each clone. |
| Clonality: | Monoclonal |
| Reactivity: | Cryptosporidium spp.; See Comments section below for details on each clone. |
| Immunogen: | MAb C4A1- or MAb C6B6- Iimmunoaffinity-purified C. parvum antigens |
| Species Immunized: | Mouse |
| Purification Method: | Not purified |
| Buffer: | Spent cell culture supernatant medium (containing DMEM, 15% FBS, pen/strep) |
| Tested Applications: | IFA, Immunoblot |
| Storage: | -80C |
| Shipped: | Dry Ice |
Data
Western Blot analysis of MAb Reactivities
Excysted oocysts were solubilized in lysis buffer containing protease inhibitors, boiled in reducing SDS-PAGE sample buffer, resolved in 4 to 20% gradient SDS-PAGE reducing gels, and Western blotted as previously described (Schaefer et. al. 2000. Infect. Immun. 68:2608-2616). Blots were probed with culture supernatants containing MAbs generated against C. parvum or isotype-matched control MAbs of irrelevant specificity (4 ug/ml for immunoglobulin M [IgM] and 10 ug/ml for IgG). Bound MAb was detected with affinity-purified phosphatase conjugated goat anti-mouse IgG, IgA, and IgM and phosphatase substrate. Molecular mass standards are indicated on the left (myosin, 208 kDa; b-galactosidase, 144 kDa; BSA, 87 kDa; carbonic anhydrase, 44.1 kDa; soybean trypsin inhibitor, 32.7 kDa; lysozyme, 17.7 kDa [Bio-Rad].
Indirect Fluorescent Antibody (IFA)
Provider
From the laboratory of Michael W. Riggs, DVM, PhD, University of Arizona.
Comments
| Clone | Target | Epitope Structure1 | Isotype | Comments |
| 1E3 | GP25-200 | P/PD | IgG1 k | Binds to air dried or formalin fixed intact C. parvum oocysts and excysted oocyst shells by IFA; binds to solubilized C. hominis oocysts in dot immunoblot. |
| 2A11 | GP25-200 | pending | IgM k | Binds to the suture of air dried or formalin fixed excysted C. parvum oocyst shells by IFA; binds to solubilized C. hominis oocysts in dot immunoblot. |
| 2B4 | GP25-200 | C/CD | IgG2a k | Binds to air dried amorphous soluble antigen deposits (possibly secreted exoantigens) from excysted C. parvum sporozoite preps in IFA; binds to solubilized C. hominis oocyst antigens in western blot. |
| 2C2 | GP25-200 | P/PD | IgG1k | Binds to the suture of air dried or formalin fixed excysted C. parvum oocyst shells in IFA; binds to air dried excysted C. hominis oocyst shells in IFA and solubilized C. hominis oocysts in dot immunoblot. |
| 2F12 | GP25-200 | P/PD | IgG2a k | Binds to air dried excysted oocyst shells and air dried intact oocysts of C. parvum, C. hominis, C. meleagridis and C. muris in IFA; binds to solubilized C. hominis oocyst antigens in western blot. |
| 2G1 | GP25-200 | pending | IgG2b k | Binds to the suture of air dried excysted C. parvum and C. hominis oocyst shells and air dried intact C. parvum and C. hominis oocysts by IFA; binds to solubilized C. hominis oocysts in dot immunoblot. |
| 3H2 | GP25-200 | C/CD | IgM | Reduces C. parvum infection in the neonatal mouse model; binds to anterior surface and multifocally on air dried C. parvum and C. hominis sporozoites, intact oocysts and excysted oocyst shells in IFA; binds to live C. parvum sporozoites with posterior capping in live IFA; binds to solubilized C. hominis oocyst antigens in western blot. |
| 4D3 | GP25-200 | P/PD | IgM k | Binds to air dried or formalin fixed intact oocysts and excysted oocyst shells of C. parvum, C. muris and C. meleagridis in IFA; binds to solubilized C. hominis oocyst antigens in western blot. |
| 4D5 | GP25-200 | P/PD | IgG1 k | Binds to air dried excysted C. parvum oocyst shells in IFA; binds to solubilized C. hominis oocyst antigens in dot immunoblot. |
| 4E11 | GP25-200 | C/CD | IgM k | Binds to surface and anterior antigens of air dried or formalin fixed C. parvum sporozoites and excysted oocyst shells in IFA; binds to the surface of live C. parvum sporozoites in live IFA; binds to air dried intact C. muris and C. meleagridis oocysts in IFA; binds to solubilized C. hominis oocyst antigens in western blot. |
| 3D1 | P23 | P/PD | IgM | Binds diffusely to the surface of air dried or formalin fixed C. parvum sporozoites and to air dried soluble antigen deposited in trails during sporozoite gliding motility in IFA; binds to solubilized C. hominis oocysts in western blot; reactivity with P23 rC7 (3) pending. |
| 5B9 | P23 | pending | IgM | Reduces C. parvum infection in the neonatal mouse model; binds to air dried soluble antigen deposited in trails during sporozoite gliding motility and to the surface of air dried C. parvum sporozoites in IFA; unreactive with solubilized C. parvum oocyst antigens in western blot or dot immunoblot; unreactive with P23 rC7 (3) in dot immunoblot. |
References
- Schaefer, DA, Auerbach-Dixon, BA, and Riggs, MW. Characterization and formulation of multiple epitope-specific neutralizing monoclonal antibodies for passive immunization against cryptosporidiosis. 2000. Infect. Immun., 68(5), 2608-2616.
- Riggs, MW, Stone, AL, Yount, PA, Langer, RC, Arrowood, MJ, and Bentley, DL. Protective monoclonal antibody defines a circumsporozoite-like glycoprotein exoantigen of Cryptosporidium parvum sporozoites and merozoites. 1997. J Immunol. 158, 1787-1795.
- Perryman, LE, DP Jasmer, MW Riggs, SG Bohnet, TC McGuire and MJ Arrowood. 1996. A Cloned Gene of Cryptosporidium parvum Encodes Neutralization-Sensitive Epitopes. Mol. Biochem. Parasitol. 80:137-147.
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