Anti-P2X2 Receptor Antibody

Anti-P2X2 Receptor Antibody is an antibody against P2X2 Receptor for use in IC, IH & WB.

Immunoblotting: 1:500

Immunohistochemistry: 1:500.

Immunocytochemistry: 1:500.

Optimal working dilutions must be determined by end user.

APPLICATION NOTES FOR AB5894

IMMUNOHISTOCHEMISTRY

Male Sprague-Dawley rats (b.wt. 100-150g) were anesthetized with sodium pentobarbital and perfused via the ascending aorta with: 1) 50 mL 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 0.1 M phosphate buffer (pH 7.4) containing 10% sucrose. Slide-mounted tissue sections were incubated with blocking buffer for 1 hour at room temperature. Primary antibody was diluted in blocking buffer to the appropriate working dilution. Blocking buffer was removed and the slides were then incubated at 2-8°C for 18-24 hours with AB5894 (1:500). After rinsing in PBS 3 times sections were incubated for 60 minutes at room temperature 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.

IMMUNOCYTOCHEMISTRYP

2X2 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 above

WESTERN BLOTTING

Cell membrane extracts were examined by electrophoresis (8% acrylamide) with SDS under reducing conditions and transferred to a nylon membrane. Membranes were blocked for 1 hour at 2-8°C with 0.1% Tween 20 and 2.5% milk powder (w/v) in PBS. Membranes were incubated with AB5894 diluted 1:500 with same buffer overnight at 2-8°C. Membranes were rinsed and incubated with HRP conjugated secondary antibody for 1 hour at room temperature. Following rinsing the membranes were processed using enhanced chemiluminescence.

Research Category
Neuroscience

Research Sub Category
Neurotransmitters & Receptors

P2X2 Receptor.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

A 13 amino acid peptide corresponding to amino acids 460-472 from the carboxy-terminus of the rat P2X2 Receptor protein.

Control peptide: Catalog Number AG354

Serum. Liquid. Contains 0.05% sodium azide.

Maintain at -20°C in undiluted aliquots for up to 6 months. Avoid repeated freeze/thaw cycles.

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany