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:
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-633Antibody (#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.
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