Anti-P2X2 Receptor

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, transferred to a nylon membrane. Membranes were blocked for 1 hour at 4?C with 0.1% Tween 20 and 2.5% milk powder in PBS. Membranes were incubated with P1003-05D (1:500 in the same buffer overnight at 4?C. Membranes were rinsed and incubated withI1904-47H: IgG, H&L (HRP) Pab GtxRt for 1 hour at RT. Following rinsing, the membranes were processed using ECL, Immunohistochemistry: 1:1000. Antiserum was used on perfusion fixed tissue. Perfusion: 1) calcium-free Tyrode's solution, 2) paraformaldehyde-picric acid 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 RT. P1003-05D 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 RT. Slides were again rinsed 3 times and coverslipped. Staining was examined using fluorescence microscopy. , Immunocytochemistry: 1:1000. P2X3 transfected cells were processed for indirect immunofluorescence. Media was removed and cells were gently washed 3 times with serum-free media. Following fixation procedure, cells were processed for indirect immunofluorescence as described above. , , 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, store 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.