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Technical Data |
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N2982-12 |
Nociceptin (Orphanin FQ) |
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Description: Applications: Suitable for use in Immunohistochemistry. Other applications not tested. Recommended Dilution: Immunohistochemistry: 1:500 This orphanin FQ robustly stains nerve fibers in brain and spinal cord. Animals were treated with colchicines to visualize orphanin FQ-expressing cell bodies. Staining was blocked by preabsorption with the cognate peptide. Antiserum was used on perfusion fixed tissue. Perfusion: 1) calcium-free Tyrode’s solution, 2) fixative, and 3) 10% sucrose in PBS as a cryo-protectant. Desired tissues were dissected and stored overnight in 10% sucrose in PBS. Slide-mounted tissue sections were processed for indirect immunofluorescence. Slides were incubated with blocking buffer for 1 hour at room temperature. Primary antiserum was diluted with blocking buffer to the appropriate working concentration. Blocking buffer was removed and slides were incubated for 18-24 hours at 4°C with primary antiserum. Slides were rinsed 3 times and then incubated with secondary antibodies for 1 hour at room temperature. Slides were again rinsed 3 times and coverslipped. Staining was examined using fluorescence microscopy. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (40-50%). Freeze at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
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