Technical Data
S8000-21
Substance P (NK, Neurokinin)
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Technical Data |
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S8000-21 |
Substance P (NK, Neurokinin) |
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Description: Applications: Suitable for use in Immunohistochemistry. Other applications not tested. Recommended Dilution: Immunohistochemistry: 1:50-1:100 Optimal dilutions to be determined by the researcher. Immunohistochemistry Protocol: Male Sprague-Dawley rats (b.wt. 100-150g) were anesthetized with sodium pentobarbital and perfused via the ascending aorta with: 1) 50ml of Ca2+ free Tyrode’s solution followed by 2) a formalin-picric acid fixative (4% paraformaldehyde with 0.4% picric acid in 0.16 M phosphate buffer, pH 6.9) and 3) 10% sucrose in PBS as a cryo-protectant. Tissues were rapidly dissected out and stored overnight in 0.1M phosphate buffer, pH 7.4 containing 10% sucrose. Slide-mounted tissue sections were incubated with blocking buffer for 1 hour at RT. Primary antibody was diluted in blocking buffer to the appropriate working dilution. Blocking buffer was removed and the slides were then incubated at 4°C for 18-24 hours. After rinsing in PBS 3 times sections were incubated for 60 minutes at RT with Cy3-conjugated secondary antibodies. After mounting in a mixture of PBS and glycerol (1:3) containing 0.1% p-phenylenediamine, sections were examined with a Nikon Microphot-SA epifluorescence 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. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. |
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