AB5477-200UL
Anti-P2X6 Receptor Antibody
Chemicon®, from rabbit
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
biological source
rabbit
antibody form
affinity purified immunoglobulin
antibody product type
primary antibodies
clone
polyclonal
purified by
affinity chromatography
species reactivity
rat
manufacturer/tradename
Chemicon®
technique(s)
western blot: suitable
NCBI accession no.
UniProt accession no.
shipped in
wet ice
target post-translational modification
unmodified
Gene Information
rat ... P2Rx6(25041)
Specificity
Recognizes P2X6 Receptor. The epitope specific for the P2X6 receptor is not present in any other known proteins.
SPECIES REACTIVITIES:
The immunogen sequence is conserved in mouse (15/15) and human (14/17). Reactivity with other species has not yet been tested.
SPECIES REACTIVITIES:
The immunogen sequence is conserved in mouse (15/15) and human (14/17). Reactivity with other species has not yet been tested.
Immunogen
Highly purified peptide corresponding to residues 363-379 of rat P2X6 (Accession P51579).
Application
Research Category
Neuroscience
Neuroscience
Research Sub Category
Neurotransmitters & Receptors
Neurotransmitters & Receptors
This Anti-P2X6 Receptor Antibody is validated for use in WB for the detection of P2X6 Receptor.
Western blotting: 1:200 using ECL on rat brain membranes.
Dilutions should be made using a carrier protein such as BSA (1-3%).
Optimal working dilutions must be determined by the end user.
SUGGESTED WESTERN BLOT PROTOCOL
1. Mix the samples (organ membranes: 50 μg/lane; transfected cells: 500,000 cells/lane) with sample-buffer X 2, and heat 10 min at 70°C.
2. 5-50 μL applied to Minigel lane (0.75-1.5 mm width) and run at standard conditions. (60 mA for 2 1.5 mm Minigel gels, 1.4 h). It is suggested that you run 5-15% acrylamide (37.5:1 acrylamide:bisacrysmide) minigel (1.5 mm width) at 30 mA/gel ~1-1.5 hours.
3. Transfer in semi-dry system under standard conditions (3 h 100 mA for two minigel gels)
4. Stain the transferred bands with Chemicon BLOT-FastStain (Catalog Number 2076).
5. Destain with deionized water.
6. Block with 5% non-fat milk (Marvel or Carnation) in PBS, and 0.025 % sodium azide, overnight at 2-8°C. The non-fat milk should be dissolved freshly, centrifuged 10,000 rpm for 10 min, and filtered through glass filter (Gelman Acrodisc).
7. Incubation with first antibody 2 h at room temperature or overnight at 4°C in blocking solution. The antibody preparation should be centrifuged before use (10,000 g 5 min.). Optimal working dilutions and incubation time will need to be determined by the end user.
8. Wash 4 x 10 min. with PBS-0.1% tween 20. From this stage, azide should be omitted.
9. Incubation with the secondary antibody (HRP-conjugated goat anti-rabbit antibody, for example Chemicon Catalog Number AP132P, diluted appropriately) 1 h at room temperature.
10. Wash 4 x 10 min. with PBS-0.1% tween 20.
11. Perform ECL with commercial kits (ChemiLUCENT, Chemicon Catalog Number 2600).
Dilutions should be made using a carrier protein such as BSA (1-3%).
Optimal working dilutions must be determined by the end user.
SUGGESTED WESTERN BLOT PROTOCOL
1. Mix the samples (organ membranes: 50 μg/lane; transfected cells: 500,000 cells/lane) with sample-buffer X 2, and heat 10 min at 70°C.
2. 5-50 μL applied to Minigel lane (0.75-1.5 mm width) and run at standard conditions. (60 mA for 2 1.5 mm Minigel gels, 1.4 h). It is suggested that you run 5-15% acrylamide (37.5:1 acrylamide:bisacrysmide) minigel (1.5 mm width) at 30 mA/gel ~1-1.5 hours.
3. Transfer in semi-dry system under standard conditions (3 h 100 mA for two minigel gels)
4. Stain the transferred bands with Chemicon BLOT-FastStain (Catalog Number 2076).
5. Destain with deionized water.
6. Block with 5% non-fat milk (Marvel or Carnation) in PBS, and 0.025 % sodium azide, overnight at 2-8°C. The non-fat milk should be dissolved freshly, centrifuged 10,000 rpm for 10 min, and filtered through glass filter (Gelman Acrodisc).
7. Incubation with first antibody 2 h at room temperature or overnight at 4°C in blocking solution. The antibody preparation should be centrifuged before use (10,000 g 5 min.). Optimal working dilutions and incubation time will need to be determined by the end user.
8. Wash 4 x 10 min. with PBS-0.1% tween 20. From this stage, azide should be omitted.
9. Incubation with the secondary antibody (HRP-conjugated goat anti-rabbit antibody, for example Chemicon Catalog Number AP132P, diluted appropriately) 1 h at room temperature.
10. Wash 4 x 10 min. with PBS-0.1% tween 20.
11. Perform ECL with commercial kits (ChemiLUCENT, Chemicon Catalog Number 2600).
Physical form
Affinity purified immunoglobulin. Lyophilized from PBS, pH 7.4, containing 1% BSA and 0.05% sodium azide. Reconstitute with 200μL of sterile distilled water. Centrifuge antibody preparation before use (10,000 x g for 5 min).
Storage and Stability
Maintain lyophilized material at -20°C for up to 6 months after date of receipt. After reconstitution maintain at -20°C in undiluted aliquots for up to 6 months. Avoid repeated freeze/thaw cycles.
Analysis Note
Control
Included free of charge with the antibody is 40 μg of control antigen (lyophilized powder). For negative control, preincubate 1 μg of purified peptide with 1 μg of antibody for one hour at room temperature. Optimal concentrations must be determined by the end user.
Included free of charge with the antibody is 40 μg of control antigen (lyophilized powder). For negative control, preincubate 1 μg of purified peptide with 1 μg of antibody for one hour at room temperature. Optimal concentrations must be determined by the end user.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
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.
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Storage Class
10 - Combustible liquids
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Juan Ignacio Díaz-Hernández et al.
PloS one, 10(4), e0123121-e0123121 (2015-04-16)
P2X receptors are ligand-gated ion channels sensitive to extracellular nucleotides formed by the assembling of three equal or different P2X subunits. In this work we report, for the first time, the accumulation of the P2X6 subunit inside the nucleus of
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