84-1050-49　［取扱停止］P2X3 Receptor 150ul　P1003-08B

特徴

• Applications:Suitable for use in Western Blot, Immunohistochemistry and Immunocytochemistry.
• Other applications not tested.
• Recommended Dilution:Western Blot (Reducing): 1:1000.
• Cell membrane extracts were examined by SDS-PAGE under reducing conditions and transferred to a nylon membrane.
• Membranes were blocked for 1 hour at 4˚C with PBS, 0.1% Tween 20, 2.5% milk powder.
• Membranes were incubated with diluted P1003-08B overnight at 4˚C then rinsed with PBS and incubated with IgG (HRP) Goat anti-rat for 1 hour at RT.
• Following rinsing, the membranes were processed using ECLImmunohistochemistry: 1:1000.
• Antiserum was used on perfusion fixed tissue.
• Perfusion: 1) calcium-free Tyrode’s solution.
• 2) Paraformaldehyde-picric acid fixative.
• 3) PBS, 10% sucrose as a cryo-protectant.
• Desired tissues were dissected and stored overnight in PBS, 10% sucrose.
• Slide-mounted tissue sections were processed for Indirect Immunofluorescence.
• Slides were incubated with blocking buffer for 1 hour at RT.
• P1003-08B was diluted with blocking buffer to the appropriate working concentration.
• Blocking buffer was removed.
• Slides were incubated for 18-24 hours at 4°C with primary antiserum.
• Slides were rinsed 3 times with PBS.
• Slides were incubated with secondary antibodies for 1 hour at RT.
• Slides were again rinsed 3 times with PBS and coverslipped.
• Staining was examined using Fluorescence Microscopy
• Immunocytochemistry: 1:1000.
• P2X3 transfected cells were processed for Indirect Immunofluorescence.
• Media was removed.
• Cells were gently washed 3 times with serum-free media.
• Following fixation procedure, cells were processed for Indirect Immunofluorescence as described for Immunohistochemistry.
• Optimal dilutions to be determined by the researcher.
• 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.