Name: Anti-P2X7 Receptor (extracellular)-ATTO Fluor-633 Antibody
Brand: Alomone Labs
SKU: APR-008-FR

Overview

Cat #: APR-008-FR

Alternative Name Purinergic receptor P2X7, P2RX7, P2Z, ATP receptor

Lyophilized Powder yes

Type: Polyclonal

Host: Rabbit

Reactivity: h, m, r

Immunogen

Peptide KKGWMDPQSKGIQTGRC, corresponding to amino acid residues 136-152 of mouse P2X7 receptor (Accession Q9Z1M0). Extracellular loop.

Accession (Uniprot) Number Q9Z1M0

Gene ID 18439

Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.

Homology Human, rat - identical; bovine - 14/17 amino acid residues identical.

RRID AB_2341046.

Purity Affinity purified on immobilized antigen.

Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.

Isotype Rabbit IgG.

Label ATTO-633. Maximum absorption 629 nm; maximum fluorescence 657 nm. The fluorescence is excited most efficiently in the 610 - 645 nm range. This label is analogous to the dyes Alexa 647, Alexa 633 and Cy5 and can be used for direct flow cytometry (FACS) using the He:Ne laser.

Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.

Reconstitution 50 µl double distilled water (DDW).

Antibody concentration after reconstitution 1 mg/ml.

Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 × g 5 min).

Standard quality control of each lot Western blot analysis (unlabeled antibody, #APR-008), and direct flow cytometry (labeled antibody).

Applications: fc, ic, if, lci

May also work in: ih*

Direct flow cytometry

Cell surface detection of P2RX7 in intact living human THP-1 monocytic leukemia cells:
^___ Cells.
^___ Cells + Rabbit IgG isotype control-APC (#RIC-001-FR).
^___ Cells + P2X7 Receptor (extracellular)-ATTO Fluor-633 (#APR-008-FR), (5µg).

Live cell imaging / Immunocytochemistry

Expression of P2RX7 in rat brain glioma (C6) cells
Cell surface detection of P2RX7 in intact living rat C6 cells. A. Extracellular staining of cells with Anti-P2X7 Receptor (extracellular)-ATTO Fluor-633 Antibody (#APR-008-FR), (1:25, purple). B. Merged image of A and nuclear staining using DAPI as the counterstain (blue). C. Merged image of B with cells live image.

References

Ding, Y. et al. (2000) J. Auton. Nerv. Syst. 81, 289.
Kim, M. et al. (2001) EMBO J. 20, 6347.
Chizh, B.A. and Illes, P. (2001) Pharmacol. Rev. 53, 553.
Nihei, O.K. et al. (2000) Mem.Inst.Oswaldo Cruz. 95, 415.

Scientific background

The P2X7 receptor is a member of the ionotropic P2X receptor family that is activated by ATP. To date, this family is composed of seven cloned receptor subtypes, named P2X1-P2X7.

The different P2X receptors show distinct expression patterns. P2X1-6 have been found in the central and peripheral nervous system, while the P2X7 receptor is found in cells of the immune system, particularly antigen presenting cells, and microglia. The P2X7 receptor mediates the release of proinflamatory cytokines, stimulation of transcription factors and may also have an important role in apoptosis.^1-3

Different techniques have been used to characterize the P2X7 receptor. Most of them investigated pores, ion channels (electrophysiology) and membrane alterations (calcium microfluorometry, dye uptake, membrane depolarization and ion influx analysis). With the introduction of flow cytometry, it is now possible to analyze multiple cell parameters such as cell cycle, cell membrane alteration, calcium influx and cell phenotype.⁴

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat

Lyophilized Powder

Anti-P2X7 Receptor (extracellular) Antibody (#APR-008) is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot, indirect flow cytometry, immunohistochemistry, live cell imaging, and immunocytochemistry applications. It has been designed to recognize P2X7 purinergic receptor from mouse, rat, and human samples.

Anti-P2X7 Receptor (extracellular)-ATTO Fluor-633 Antibody (#APR-008-FR) is directly labeled with an ATTO-633 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. ATTO 633 has a maximum absorption at 629 nm and a maximum fluorescence at 657 nm. The fluorescence is excited most efficiently in the range 610 to 645 nm. This label is analogous to the well-known dyes Alexa 647, Alexa 633 and Cy5. Anti-P2X7 Receptor (extracellular)-ATTO Fluor-633 Antibody is especially suited for experiments requiring simultaneous labeling of different markers.

Certificate of Analysis SDS

For research purposes only, not for human use

Last Update: 22/02/2024

Applications

Citations

KO validation citations

Western blot analysis of mouse neutrophil lysate using #APR-008. Tested in P2X7^-/- mice.
Karmakar, M. et al. (2016) Nat. Commun. 7, 10555.

Specifications

Scientific Background

Specific Control Product

P2X7 Receptor (extracellular) Blocking Peptide (#BLP-PR008) is the original antigen used for immunization during Anti-P2X7 Receptor (extracellular) Antibody (#APR-008) generation. The blocking peptide binds and ‘blocks’ Anti-P2X7 Receptor (extracellular) primary antibody, this makes it a good negative reagent control to help confirm antibody specificity in western blot and immunohistochemistry applications. This control is also often called a pre-adsorption control.

P2X7 Receptor (extracellular) Blocking Peptide (#BLP-PR008)

Shipping and Ordering Information

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Anti-P2X7 Receptor (extracellular)-ATTO Fluor-633 Antibody