Description		Monoclonal antibody cocktail against glial fibrillary acidic protein (GFAP) derived from the Bigner-Eng clones MAb1B4, MAb2E1 and MAb4A11
Clone		SMI-22
Form		Ascites Fluid
Host		Mouse
Species Reactivity		Human, sheep, cow, dog, pig, rat, guinea pig, rat, mouse and chicken
IsoType		IgG2b
Specificity		The Bigner-Eng antibodies have originally been assayed by indirect radioimmunoassay against fixed cell monolayers of a GFAP-positive human glioma cell line and also by competitive radioimmunoassay with radiolabelled GFAP and by competitive immunoradioassay with radiolabelled antibody. They have been further characterized by immunoblots of GFAP and by immunocytochemistry with the peroxidase-antiperoxidase method. Each of the components of SMI 22 is specific for GFAP and visualizes immunocytochemically astrocytes and astrocytic processes as well as Bergman glia in a wide variety of species (human, sheep, cow, dog, pig, rat, guinea pig, rat, mouse and chicken). The mixture of the three antibodies provides, however, for a more comprehensive detection of astrocytomas than each antibody alone. Both, anaplastic and reactive astrocytes are stained immunocytochemically. Metastatic tumors and brain tumors of non-astrocytic origin, such as medulloblastomas, meningiomas, choroid plexus papillomas and schwannomas are not stained. There appears to be a positive correlation between fibrillarity of immunocytochemical localization in astrocytomas and their degrees of differentiation, and a negative correlation between the percentage of positive cells and the degree of anaplasticity. In morphologically diagnosed ependymomas and oligodendrogliomas positive reaction appears to reveal presence of malignant astrocytes.
Uses		This antibody is effective in immunoblotting, immunocytochemistry and ELISA.
Suggested Working Dilution		The extent of permissible dilution of SMI 22 beyond those recommended for general application depends upon nature and concentration of the antigen examined, species of the antigen, method of fixation and kind of section examined. This antibody is sold for laboratory research use only, not for human or in-vivo use. Covance antibodies may not be resold or modified for resale without prior written approval. Western blot: 1:1,000 Immunohistochemistry: 1:1,000 ELISA: 1:1,000 Tissue Preparation: SMI 22 can be used to detect GFAP in paraffin, vibratome and frozen tissue sections and in cell cultures. In order to facilitate antibody reactivity on paraffin sections, the sections (on glue-coated slides) can be incubated with 0.1% trypsin in 0.05M Tris buffer, pH 7.6, for 20 to 30 minutes at 37°C after paraffin removal. Glue-coated slides are prepared by mixing 2.25g of Elmer's glue in 15 ml distilled water. Apply one drop of glue solution to a clean glass slide and smear over the slide with another clean slide. Dry overnight in a 56°C oven. Cut paraffin sections, place on glue-covered slide and dry. Alternatively, enhanced reaction for GFAP in paraffin sections can be obtained by autoclaving deparaffinized, formalin-fixed sections in distilled water (Shin et al, Lab Invest, 64:693, 1991) or by boiling sections or tissue blocks immersed in tris buffered saline, pH 9.0, in a microwave oven for 15 min (Evers and Uylings, J Neurosci Methods 72:197, 1997). Post-fixation for 10 minutes in cold methanol or methanol/hydrogen peroxide is necessary for good access of SMI 22 to astrocytes in culture or in cryostat and vibratome tissue sections.
Storage		Store at -20°C. Upon initial thawing, apportion into working aliquots and store at -20°C. Avoid repeated freeze-thaw cycles to prevent denaturing the antibody.
References		Vick WW, Bigner SH, Wikstrand CJ, Bullard DE, Kemshead J, Coakham HB, Schlom J, Johnston WW, Bigner DD: The use of a panel of monoclonal antibodies in the evaluation of cytologic specimens from the central nervous system. Acta. Cytol. 31:816, 1987. McLendon RE, Burger PC, Pegram CN, Eng LP, Bigner DD: The immunohistochemical application of three anti-GFAP monoclonal antibodies to formalin-fixed, paraffin-embedded, normal and neoplastic brain tissues. J. Neuropath. Exp. Neurol. 45:692, 1986. Pegram CN, Eng LF, Wikstrand CJ, McComb RD, Lee Y-L, Bigner DD: Monoclonal antibodies reactive with epitopes restricted to glial fibrillary acidic proteins of several species. Neurochem. Path. 8:119, 1985.
Warranty/Conditions		Covance products may not be resold or modified for resale without prior written approval.
Rev. Date		6/7/2005