Suitable for use in Immunohistochemistry, Dot Blot and Immunoprecipitation. Not recommended for Western Blot. Other applications have not been tested.
Recommended Dilutions
Immunohistochemistry (unfixed, frozen): 10ug/ml using monkey brain sections Dot Blot: 0.5-2ug/ml detects mouse Laminin in a dot blot assay. Optimal dilutions to be determined by the researcher.
Hybridoma
Mouse-rat hybridoma of X63.Ag8 653 mouse myeloma with Fisher rat splenocytes.
Immunohistochemistry 1. 10um unfixed, frozen tissue sections were washed with PBS twice, for 20 minutes. 2. Add 400ul of 8% albumin in PBS and incubate for 1 hour at room temperature. 3. Wash the sections with PBS four times, 15 minutes each, at room temperature. 4. Incubate the sections with a 10ug/ml a-Mouse Laminin B2 Chain in 1.0% albumin in PBS and incubate overnight at 4°C. Also run a negative control (no primary antibody) to check for non-specific staining. 5. Wash the sections with PBS twice for 30 minutes. 6. Incubate the sections with a 1:100 dilution of goat anti-rat IgG fluorescein conjugated secondary antibody in 1% BSA in PBS for 2 hours at room temperature. 7. Wash the section with PBS three times, 15 minutes each. 8. Examine the sections under a fluorescent microscope.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Murine EHS laminin.
Form
Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide.
Purity
Purified by Protein G affinity chromatography from cell culture.
Specificity
Recognizes and is specific for the mouse B2 chain laminin. Does not crossreact with other basement membrane components or fibronectin. Species Crossreactivity: human and monkey
References
1. Ljubimov et al., Exp. Cell Res. 165: 530, 1986.