P2X3 Receptor (P2RX3) Antibody – Rabbit Polyclonal

Suitable for use in Western Blot, Immunohistochemistry and Immunocytochemistry.
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 ECL
Immunohistochemistry: 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 degree 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.