Description		Monoclonal antibody against glial fibrillary acidic protein (GFAP) derived from the Bigner-Eng clone MAb 4A11
Clone		SMI-25
Form		Ascites Fluid
Host		Mouse
Species Reactivity		Human
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. While SMI 25 provides for general visualization of GFAP in astrocytes and Bergman glia, it detects a more selected group of astrocytomas than mixtures of SMI 23, 24 and 25. The significance of this selectivity has not yet been explored. Possibilities are different characteristics of individual tumors or cooperativity of antibodies.
Uses		This antibody is effective in immunoblotting, immunocytochemistry and ELISA.
Suggested Working Dilution		The extent of permissible dilution of SMI 25 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 25 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 37oC 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 56oC oven. Cut paraffin sections, place on glue-covered slides and dry. Alternatively, enhanced reaction for GFAP in paraffin sections can be obtained by autoclaving deparaffinized, formalin-fixed sections in distilled water as described by 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 in methanol or methanol/hydrogen peroxide is necessary for good access of SMI 25 to astrocytes in culture or in cryostat and vibratome sections of tissue perfused with 4% paraformaldehyde.
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.
Warranty/Conditions		Covance products may not be resold or modified for resale without prior written approval.
Rev. Date		6/7/2005