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Applikationshinweise
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Paraffin Sections Anti-calponin, CALP can be used on formalin-fixed, paraffin-embedded tissue sections. Pretreatment of tissue with proteolytic enzymes should be performed prior to staining. For improved staining results, the deparaffinized tissue sections may be treated with a proteolytic enzyme followed by target retrieval. For greater adherence of tissue sections to glass slides, the use of Silanized Slides (code S3003) is recommended. Deparaffinized tissue sections should first be treated for 5 to 10 minutes with a mild enzyme solution. A recommended proteolytic enzyme is Proteinase K (code S3004) which must be further diluted 1:500 in 0.05 mol/L Tris-HCl, pH 7.6 to give a final concentration of 0.04 mg/mL. (106948-002) 303341EFG_001 p. 2/5 Following proteolytic digestion, tissue sections can be treated with heat. When using the water bath method, preheat a Coplin jar containing 0.01 mol/L citrate buffer, pH 6.0 as well as a water bath to 95-99 °C. When the temperature has stabilized, place tissue sections into the Coplin jar containing the preheated buffer. Heat the tissue sections for 40 minutes. For improved staining results and a shorter incubation time, Target Retrieval Solution (code S1700) can be used in place of the 0.01 mol/L citrate buffer. Under these conditions the incubation time in the water bath may be reduced to 20 minutes. After thermal treatment, allow the jar with buffer and slides to cool for 20 minutes at room temperature. Rinse well with distilled water and place slides into buffer. Cryostat Sections and Cell Smears Anti-calponin, CALP can also be used to label cryostat sections or cell smears. Follow the recommended procedure for the detection system selected. Staining interpretation The staining pattern for anti-calponin is cytoplasmic. Normal Cells In immunohistochemical (IHC) studies on cryostat sections of human fetal tissue, monoclonal anti-calponin, CALP, was found to stain developing visceral smooth muscle of trachea, jejenum, esophagus and uterus in 10 and 20 week-old fetuses. Monoclonal anti-calponin did not react positively with 10 and 20 week-old fetal aortic smooth muscle cells. 1 Monoclonal antibody CALP was found to localize calponin in cryostat sections of adult visceral and vascular smooth muscle but not in epithelial cells, endothelial cells, or connective tissue fibroblasts. Adult aortic cells of the tunica media and a portion of subendothelial intimal cells were found to stain positively. 1 In cryostat sections and routinely fixed specimens of normal human breast, calponin expression has been demonstrated in smooth muscle cells of blood vessels and myoepithelial cells in the lobules, ducts and galactophorous sinuses. 6,7 Periacinar and periductal myoepithelial cells of the salivary gland have also been shown to react positively with anti-calponin, whereas ductal epithelial cells were negative. 8 Tumor Cells Calponin expression has been demonstrated by IHC in myoepithelial cells in benign and malignant breast lesions. 6,7 Myoepithelial cells in papillomas of the breast were found to stain positively with anti-calponin but no reactivity was observed in intracystic papillary carcinomas. In ductal carcinoma in situ, calponin immuno-reactivity has been demonstrated in myoepithelial cells at the periphery of involved ducts and lobules in complex sclerosing lesions of the breast. 7 Anti-calponin was shown to label stromal myofibroblasts in the majority of invasive breast carcinomas tested but was unreactive with tumor cells. 6,7 Reactivity has also been observed in neoplastic myoepithelium of routinely fixed pleomorphic adenomas of the salivary gland.
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Lagerung
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Store at 2-8 °C. Precautions: 1. For professional users. 2. This product contains sodium azide (NaN 3 ), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, NaN 3 may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used. 4. Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin. 5. Unused reagents should be disposed of according to local, State, and Federal regulations.
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Alternativen 
